Compositions and methods for treating autism

ABSTRACT

The invention provides compositions and methods of treating autism. Specifically, the invention relates to treating the core symptoms of autism by administering α-methyl-DL-tyrosine.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part (CIP) application of U.S.patent application Ser. No. 15/695,327, filed Sep. 5, 2017, which is acontinuation of U.S. patent application Ser. No. 15/138,733, filed Apr.26, 2016, now issued as U.S. Pat. No. 9,757,348, which is a continuationof U.S. patent application Ser. No. 14/520,116, filed Oct. 21, 2014, nowissued as U.S. Pat. No. 9,326,962, which is a continuation-in-part ofU.S. patent application Ser. No. 14/062,165, filed Oct. 24, 2013, nowabandoned, which claims priority to and the benefit of U.S. ProvisionalPatent Application No. 61/894,261, filed Oct. 22, 2013, all of which areincorporated by reference herein in their entirety.

FIELD OF THE INVENTION

The invention relates to compositions and methods of treating autism.Specifically, the invention relates to treating the core symptoms ofautism by administering α-methyl-DL-tyrosine.

BACKGROUND OF THE INVENTION

Autism spectrum disorder (ASD) or autism is defined in the Diagnosticand Statistics Manual of Mental Disorders V (DSM-5) by “difficulties insocial communication and social interaction, and restricted andrepetitive behavior, interests or activities”. Childhood autism is moreprevalent than childhood cancer, juvenile diabetes, and pediatricacquired immunodeficiency syndrome (AIDS) combined, with an estimatedprevalence of 1.5 million in the United States (US), 3 million childrenin Europe and tens of millions throughout the rest of the world. Autismalso represents a substantial economic burden in both children andadults. More disturbing is that, for no explicable reason, childhoodautism appears to be increasing at a rate of 10-17% per year. Lastly,there are currently no approved medications that address the coresymptoms of autism. Autism is a serious disease that represents an areaof significant economic burden and unmet medical need.

Autism was first described in 1943 by Dr. Leo Kanner who described his 5year old patient as, “ . . . happiest when left alone, almost nevercried to go with his mother, did not seem to notice his father'shomecomings, and was indifferent to visiting relatives . . . wanderedabout smiling, making stereotyped movements with his fingers . . . spunwith great pleasure anything he could seize upon to spin . . . . Wordsto him had a specifically literal, inflexible meaning . . . . When takeninto a room, he completely disregarded the people and instantly went forobjects”, as cited by Lai, M. C., et al. Autism. Lancet, 2014. 383(9920): p. 896-910.

Originally autism was thought to be a form of childhood schizophrenia.However, in the mid-1980s, it was found that autism is a heritabledisorder and believed to have a genetic etiology. Currently, autism isdefined diagnostically in the Diagnostic and Statistical Manual ofMental Disorders, 5^(th) Edition (DSM-5) (2013) by “difficulties insocial communication and social interaction, and restricted andrepetitive behavior, interests or activities.” The diagnostic criteriain DSM-5 for the features “difficulties in social communication andsocial interaction” of ASD include (1) deficits in social-emotionalreciprocity, ranging, for example, from abnormal social approach andfailure of normal back-and-forth conversation; to reduced sharing ofinterests, emotions, or affect; to failure to initiate or respond tosocial interactions; (2) Deficits in nonverbal communicative behaviorsused for social interaction, ranging, for example, from poorlyintegrated verbal and nonverbal communication; to abnormalities in eyecontact and body language or deficits in understanding and use ofgestures; to a total lack of facial expressions and nonverbalcommunication; and (3) deficits in developing, maintaining, andunderstanding relationships, ranging, for example, from difficultiesadjusting behavior to suit various social contexts; to difficulties insharing imaginative play or in making friends; to absence of interest inpeers.

Typically displayed in early childhood, autism is associated with manyco-morbidities that include epilepsy, Fragile-X syndrome, Rettssyndrome, attention deficit/hyperactivity disorder (ADHD), abnormalsensory or motor responses, disturbed sleep, reduced cognitivefunctionality, anxiety and aggression.

In 2010, the CDC reported the rate of autism in the US to be 1 in 68children, with boys being 5 times more susceptible than girls. Thismeans that 1 in 42 boys are diagnosed with autism. There was an increaseof about 30% since the assessment of autism prevalence conducted in2014, and more than double the rate that was reported only 12 years andthe problem is growing rapidly. In New Jersey the observed rate ofautism was 1 in 46 children, which means that 1 in 29 boys born in NewJersey are likely to be autistic.

The financial cost of raising an autistic child was estimated in 2014 tobe $3.2M more than the cost of raising a non-autistic child, and thisdoes not take into account the societal costs of maintaining thispopulation as adults once their families are no longer able to do so.The societal costs of autism are broad and deep, and many have neverbeen explored. For example, it was only in mid-2017 that information wasdeveloped on the rate of healthcare utilization by autistics and it wasfound that their need for psychiatric care as well as care for the highincidence of autism associated comorbidities was far beyond that of thegeneral population or other elements of the psychiatric patientpopulation. Similarly, it was not until September of 2017 that the rateof school suspension and expulsion was dramatically higher in theautistic population, and growing as the autistic population grew innumbers.

Recently, attention has been brought to bear on autism associatedmortality rates. Although autism is not typically considered to be afatal disease, a number of investigators have reported a significantlyincreased mortality in the autistic population with the major cause ofdeath being suicide. A matched case cohort study based upon the SwedishNational Patient Registry and the Cause of Death Registry looked atdeaths between 1987 and 2009, and found a 256% greater death rate inautistic patients compared to the general population. The mean age atthe time of death was 70.2 years for the general population and 58.39for patients with autism, with suicide associated with better performingpatients. A review of 1,706 children and adolescents reported an 18%increased risk of suicidal ideation or attempts in autism. 35% ofpatients with Asperger's syndrome were reported in a Canadian study tohave attempted suicide. Similarly, in Japan, Australia, England, andBelgium. In a French review of the PubMed literature, it was found thatoverall 21.3% of autism patients reported suicidal ideation or hadattempted suicide, with the noteworthy observation that “ . . . themethods used are often violent”.

Enormous resources are being spent on research into the causes ofautism. Genetic, dietary, developmental, pharmacologic, environmental,and behavioral elements have all been implicated as potential causes ofthis syndrome.

A purified, single enantiomer presentation ofmolecule—α-methyl-L-tyrosine, currently marketed as Demser®, is known toinhibit tyrosine hydroxylase, which is the rate limiting enzyme in thebiosynthesis of catecholamine's consequent to enzyme-mediated conversionof L-tyrosine to DOPA (dihydroxy-phenylalanine). This first step incatecholamine synthesis is highly stereospecific for L-tyrosine. L-AMPT,a methylated L-tyrosine, acts as a competitive inhibitor for tyrosinehydroxylase (Demser® Prescribing Information, 2015; FDA Summary Basis ofApproval [SBA] for Demser®, 1979). Demser® was initially approved by theFDA in 1979 and is indicated for use in the treatment of patients withpheochromocytoma for (1) the preoperative preparation of patients forsurgery, (2) management of patients when surgery is contraindicated, and(3) chronic treatment of patients with malignant pheochromocytoma. Therecommended initial dosage of Demser® for adults and children 12 yearsof age and older is 250 mg orally four times daily. This dosage may beincreased by 250 mg to 500 mg every day to a maximum of 4.0 g/day in 4divided doses. Although Demser® is commercially available, thoseordinarily skilled in the art have not developed methods for treatingautism or its core symptoms by administering a racemic mixture ofα-methyl-DL-tyrosine, i.e., DL-AMPT.

Given the widespread concern over (a) dramatic increases in prevalenceof autism, (b) devastating impact of the disorder on children and theirfamilies, and (c) substantial social and economic burden that autismplace on communities, educational, medical and mental health systemswithin our society, autism represents a public health emergency.Accordingly, there exists a vital need for therapeutically effectivemethods for treating autism, in particular, the core symptoms of thisdisorder.

SUMMARY OF THE INVENTION

In one aspect, the invention provides a method for treating an autism ina subject in need thereof, the method comprising administering to saidsubject a composition comprising a therapeutically effective amount ofα-methyl-DL-tyrosine and a pharmaceutically acceptable carrier. In anexemplary embodiment, the composition comprises α-methyl-DL-tyrosine inan amount ranging from about 50 mg (w/w) to about 500 mg (w/w).

In another aspect, the invention provides a method for providing aplasma concentration of a therapeutic drug for a long term for treatingan autism in a subject in need thereof, the method comprisingadministering to said subject said therapeutic drug at a concentrationranging from about 50 mg (w/w) to about 500 mg (w/w) three times a day,wherein said therapeutic drug is α-methyl-DL-tyrosine, wherein saidplasma concentration ranges from about 500 ng/ml to 5000 ng/ml, andwherein said term is at least 1 week.

In another aspect, the invention provides a method for treating anautism associated clinical trait in a subject in need thereof, themethod comprising administering to said subject a composition comprisinga therapeutically effective amount of α-methyl-DL-tyrosine, therebytreating said autism associated clinical trait in said subject. In anexemplary embodiment, the clinical trait is a deficit in socialcommunication, a deficit in social interaction, a deficit in socialmotivation, lethargy and social withdrawal, inappropriate speech,hyperactivity, stereotypic behavior, irritability and agitation,restrictive behavior, repetitive behavior, ritualistic behavior,sameness behavior, compulsive behavior, self-injurious behavior or acombination thereof.

Other features and advantages of the present invention will becomeapparent from the following detailed description examples and figures.It should be understood, however, that the detailed description and thespecific examples while indicating preferred embodiments of theinvention are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts Conners Parent Rating Scale Data results.

FIG. 2 illustrates patient enrollment and disposition.

FIG. 3 shows an example of largest change from baseline in vital signsnoted in a patient in Study HT 02-121 (Example 6) and demonstrates thatthese changes were transient, falling far short of ortho staticcriteria.

FIG. 4 shows random L1-79 plasma concentrations (except Day 1*) by week,overall mean and by patient from open-label 100 mg and 200 mg TID inStudy HT 02-121. *1 On Day 1, L-79 plasma concentrations were evaluatedat 1-hour post-dose.

FIG. 5 shows random L1-79 plasma concentrations by week (excluding Day1), overall mean and by patient from open-label 100 mg TID group inStudy HT 02-121.

FIG. 6 shows random L1-79 plasma concentrations by week (excluding Day1), overall Mean and by patient from open-label 200 mg TID group inStudy HT 02-121.

FIG. 7 graphically displays shows the CGI—Overall Severity (CGI-S) forL1-79 compared to placebo over time.

FIG. 8 graphically displays the CGI—Overall Improvement (CGI-I) forL1-79 compared to placebo over time.

FIG. 9 shows the change from baseline in CGI-S at week 4 (ITTPopulation, Draft).

FIG. 10 shows the change in CGI-S from Baseline to Week 4 by Patient(ITT Population, Draft).

FIG. 11 shows the responder analysis for CGI-S at Week 4/LOCF (ITTPopulation, Draft).

FIG. 12 shows the change from Baseline in VABS II StandardizedSocialization Score, Week 0-Week 8 (ITT Population, Draft).

FIG. 13 shows the change from Baseline in VABS II StandardizedCommunication Score, Week 0-Week 8 (ITT Population, Draft).

FIG. 14 shows the change from Screening in ADOS-2 Total score, Week0-Week 4 (ITT Population, Draft).

FIG. 15 shows the change from Screening in ADOS-2 Restrictive andRepetitive Behavior Total score, Week 0-Week 4 (ITT Population, Draft).

FIG. 16 shows the change from Screening in ADOS-2 Social Affect TotalScore, Week 0-Week 4 (ITT Population, Draft).

FIG. 17 shows the change in ADOS-2 Total Score from Screening to Week 4by Patient (ITT Population, Draft).

FIG. 18 shows the change from Baseline in SRS-2 Total T-score, Week0-Week 8 (ITT Population, Draft).

FIG. 19 shows the change from Baseline in SRS-2 DSM-5 SocialCommunication and Interaction T-score, Week 0-Week 8 (ITT Population,Draft).

FIG. 20 shows the change from Baseline in SRS-2 Social CommunicationT-score, Week 0-Week 8 (ITT Population, Draft).

FIG. 21 shows the change from Baseline in SRS-2 Social MotivationT-score, Week 0-Week 8 (ITT Population, Draft).

FIG. 22 shows the change from Baseline in SRS-2 DSM-5 Restrictive andRepetitive Behavior, Week 0-Week 8 (ITT Population, Draft).

FIG. 23 shows the change in SRS-2 Total T-score from Baseline to Week 4by Patient (ITT Population, Draft).

FIG. 24 shows the change in SRS-2 DSM-5 Social Communication andInteraction T-score from Baseline to Week 4 by Patient (ITT Population,Draft).

FIG. 25 shows the change in SRS-2 Social Communication T-score fromBaseline to Week 4 by Patient (ITT Population, Draft).

FIG. 26 shows the change in SRS-2 Social Motivation T-score fromBaseline to Week 4 by Patient (ITT Population, Draft).

FIG. 27 shows the responder analysis for SRS-2 Total T-score at Week4/LOCF (ITT Population, Draft).

FIG. 28 shows the responder analysis for SRS-2 DSM-5 SocialCommunication and Interaction T-score at Week 4/LOCF (ITT Population,Draft).

FIG. 29 shows the responder analysis for SRS-2 Social CommunicationT-score at Week 4/LOCF (ITT Population, Draft).

FIG. 30 shows the responder analysis for SRS-2 Social Motivation T-scoreat Week 4/LOCF (ITT Population, Draft).

FIG. 31 shows the change from Baseline in ABC-C Lethargy and SocialWithdrawal Domain, Week 0-Week 8 (ITT Population, Draft).

FIG. 32 shows the change from Baseline in ABC-C Inappropriate SpeechDomain, Week 0-Week 8 (ITT Population, Draft).

FIG. 33 shows the change from Baseline in ABC-C Hyperactivity andNoncompliance Domain, Week 0-Week 8 (ITT Population, Draft).

FIG. 34 shows the change from Baseline in ABC-C Stereotypic BehaviorDomain, Week 0-Week 8 (ITT Population, Draft).

FIG. 35 shows the change from Baseline in ABC-C Irritability andAgitation Domain, Week 0-Week 8 (ITT Population, Draft).

FIG. 36 shows the change in ABC-C Lethargy and Social Withdrawal Domainfrom Baseline to Week 4 by Patient (ITT Population, Draft).

FIG. 37 shows the change in ABC-C Inappropriate Speech Domain fromBaseline to Week 4 by Patient (ITT Population, Draft).

FIG. 38 shows the responder analysis for ABC-C Lethargy and SocialWithdrawal Domain at Week 4/LOCF (ITT Population, Draft).

FIG. 39 shows the responder Analysis for ABC-C Inappropriate SpeechDomain at Week 4/LOCF (ITT Population, Draft).

FIG. 40 shows the change from Baseline in RBS-R Total Score, Week 0-Week8 (ITT Population, Draft).

FIG. 41 shows the change from Baseline in RBS-R Restrictive Behavior,Week 0-Week 8 (ITT Population, Draft). There were mean improvements inRBS-R Total Score as noted by a decrease in score for L1-79 200 mgcompared to placebo.

FIG. 42 shows the change from Baseline in RBS-R Ritualistic Behavior,Week 0-Week 8 (ITT Population, Draft).

FIG. 43 shows the change from Baseline in RBS-R Sameness Behavior, Week0-Week 8 (ITT Population, Draft).

FIG. 44 shows the change from Baseline in RBS-R Compulsive Behavior,Week 0-Week 8 (ITT Population, Draft).

FIG. 45 shows the change from Baseline in RBS-R Stereotypic Behavior,Week 0-Week 8 (ITT Population, Draft).

FIG. 46 shows the change from Baseline in RBS-R Self-injurious Behavior,Week 0-Week 8 (ITT Population, Draft).

FIG. 47 shows the change in RBS-R Total Score from Baseline to Week 4 byPatient (ITT Population, Draft)

FIG. 48 shows the responder analyses (defined as an improvement or noworsening) for RBS-R Total Score at Week 4 or LOCF.

DETAILED DESCRIPTION OF THE INVENTION

The present subject matter may be understood more readily by referenceto the following detailed description which forms a part of thisdisclosure. It is to be understood that this invention is not limited tothe specific products, methods, conditions or parameters describedand/or shown herein, and that the terminology used herein is for thepurpose of describing particular embodiments by way of example only andis not intended to be limiting of the claimed invention.

Unless otherwise defined herein, scientific and technical terms used inconnection with the present application shall have the meanings that arecommonly understood by those of ordinary skill in the art. Further,unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular.

As employed above and throughout the disclosure, the following terms andabbreviations, unless otherwise indicated, shall be understood to havethe following meanings.

In the present disclosure the singular forms “a,” “an,” and “the”include the plural reference, and reference to a particular numericalvalue includes at least that particular value, unless the contextclearly indicates otherwise. Thus, for example, a reference to “acompound” is a reference to one or more of such compounds andequivalents thereof known to those skilled in the art, and so forth. Theterm “plurality”, as used herein, means more than one. When a range ofvalues is expressed, another embodiment includes from the one particularand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it isunderstood that the particular value forms another embodiment. Allranges are inclusive and combinable.

As used herein, the terms “component,” “composition,” “composition ofcompounds,” “compound,” “drug,” “pharmacologically active agent,”“active agent,” “therapeutic,” “therapy,” “treatment,” or “medicament”are used interchangeably herein to refer to a compound or compounds orcomposition of matter which, when administered to a subject (human oranimal) induces a desired pharmacological and/or physiologic effect bylocal and/or systemic action.

As used herein, the ensuing terms are defined, as follows:

-   ABA Applied Behavioral Analysis-   ABC-C Aberrant Behavior Checklist-Community-   ADHD Attention Deficit Hyperactivity Disorder-   ADIR Autistic Diagnosis Interview Review-   ADME Absorption, Distribution, Metabolism and Elimination-   ADOS-2 Autism Diagnosis Observation Schedule, Second Edition-   AE Adverse Events-   AIDS Acquired immunodeficiency syndrome-   ASD Autism spectrum disorder-   AST Aspartate Aminotransferase-   ALT Alanine Aminotransferase-   AUC Area under the concentration time curve-   AUC₀₋₈ Area under the concentration time curve from 0 to 8 hours-   AUC₀₋₁₂ Area under the concentration time curve from 0 to 12 hours-   AUC_(0-∞) Area under the concentration time curve from 0 to infinity-   AUC_(0-tz) Area under the concentration time curve from 0 to the    last quantifiable concentration-   CFR Code of Federal Regulations-   CGI-I Clinical Global Impression-Overall Improvement-   CGI-S Clinical Global Impression-Severity of Illness-   Cmax Maximum Concentration-   CNS Central Nervous System-   DL-AMPT Racemic (D and L isomer) α-methyl-para-tyrosine-   DOPA Dihydroxy-phenylalanine-   DSM-5 Diagnostic and Statistics Manual of Mental Disorders V-   EI Early Intervention-   EOP2 End of Phase 2-   FDA Food and Drug Administration-   FDASIA Food and Drug Administration Safety and Innovation Act-   FOBs Functional Observation Battery Assessments-   GMP Good Manufacturing Practice-   HBV Hepatitis B Virus-   HCVA Hemorrhagic Cerebrovascular Accident-   HED Human Equivalent Doses-   HIV Human Immunodeficiency Virus-   IEP Individualized Educational Program-   L-AMPT L isomer of α-methyl-para-tyrosine-   LC-MS/MS Liquid Chromatographic Mass Spectrometric-   LOCF Last Observation Carried Forward-   LQTS Long QT Syndrome-   MTD Maximum Tolerated Dose-   NDA New Drug Application-   NOAEL No Observed Adverse Effect Level-   PD Pharmacodynamic-   PK Pharmacokinetic-   PND Post Natal Day-   PSP Pediatric Study Plan-   RBS-R Repetitive Behavior Score-Revised-   SBA Summary Basis of Approval-   SRS-2 Social Responsiveness Scale, Second Edition-   t_(1/2) Half Life-   TdP Torsade's de Pointes-   TID Three times daily-   TK Toxicokinetic-   TQT Thorough QT-   ULN Upper Limit of Normal-   USAN United States Adopted Name-   VABS II Vineland Adaptive Behavior Scales, Second Edition

With respect to autism, for example, the negative effect or symptoms caninclude any of those that are the subject of the diagnostic criteriaspecified for autism spectrum disorder in the American PsychiatricAssociation's Diagnostic and Statistical Manual, Fifth Edition (DSM-5,DSM-V), the contents of which are incorporated by reference herein,e.g., deficits in social-emotional reciprocity (ranging, for example,from abnormal social approach and failure of normal back-and-forthconversation; to reduced sharing of interests, emotions, or affect; tofailure to initiate or respond to social interactions) deficits innonverbal communicative behaviors used for social interaction (ranging,for example, from poorly integrated verbal and nonverbal communication;to abnormalities in eye contact and body language or deficits inunderstanding and use of gestures; to a total lack of facial expressionsand nonverbal communication); deficits in developing, maintaining, andunderstanding relationships (ranging, for example, from difficultiesadjusting behavior to suit various social contexts; to difficulties insharing imaginative play or in making friends; to absence of interest inpeers); stereotyped or repetitive motor movements, use of objects, orspeech (e.g., simple motor stereotypes, lining up toys or flippingobjects, echolalia, idiosyncratic phrases); insistence on sameness,inflexible adherence to routines, or ritualized patterns of verbal ornonverbal behavior (e.g., extreme distress at small changes,difficulties with transitions, rigid thinking patterns, greetingrituals, need to take same route or eat same food every day); highlyrestricted, fixated interests that are abnormal in intensity or focus(e.g., strong attachment to or preoccupation with unusual objects,excessively circumscribed or perseverative interests); and hyper- orhyporeactivity to sensory input or unusual interest in sensory aspectsof the environment (e.g. apparent indifference to pain/temperature,adverse response to specific sounds or textures, excessive smelling ortouching of objects, visual fascination with lights or movement).

Some subpopulations of patients on the autism spectrum, include thosepatients diagnosed with Asperger's Disorder (i.e., Asperger Syndrome) orSocial Communication Disorder, exhibit symptoms of Attention DeficitHyperactivity Disorder (ADHD) (e.g., inattention, hyperactivity, andimpulsivity) and/or tics (motor tics or vocal tics). See, e.g., DSM-5.

Assessment of autism symptoms, or any of the symptoms of the presentdisclosure, can be performed using methods known in the art. Forexample, one method of assessing autism symptoms is the Clinical GlobalImpressions (CGI) scale based upon changes from baseline in variouspsychometric tests. These tests can include the Aberrant BehaviorChecklist-Community (ABC-C), Conners Parent Rating Scale and the AutismDiagnostic Observation Schedule (ADOS). Autism symptoms can also beassessed from the clinician's personal, clinical observations, fromvideographic information taken at regularly scheduled clinic visits, andfrom information provided by the subject's caregivers over time.Compositions and methods of the disclosure result in a reduction of atleast 1 point in at least one dimension of the Conners Parent RatingScale assessment score.

As employed above and throughout the disclosure the term “effectiveamount” refers to an amount effective, at dosages, and for periods oftime necessary, to achieve the desired result with respect to thetreatment of the relevant disorder, condition, or side effect. It willbe appreciated that the effective amount of components of the presentinvention will vary from patient to patient not only with respect to theparticular compound, component or composition selected, the route ofadministration, and the ability of the components to elicit a desiredresult in the individual, but also with respect to factors such as thedisease state or severity of the condition to be alleviated, hormonelevels, age, sex, weight of the individual, the state of being of thepatient, and the severity of the pathological condition being treated,concurrent medication or special diets then being followed by theparticular patient, and other factors which those skilled in the artwill recognize, with the appropriate dosage being at the discretion ofthe attending physician. Dosage regimes may be adjusted to provideimproved therapeutic response. An effective amount is also one in whichany toxic or detrimental effects of the components are outweighed by thetherapeutically beneficial effects.

The invention also provides a pharmaceutical composition comprisingcompounds of the invention and one or more pharmaceutically acceptablecarriers. “Pharmaceutically acceptable carriers” include any excipientwhich is nontoxic to the cell or mammal being exposed thereto at thedosages and concentrations employed. The pharmaceutical composition mayinclude one or additional therapeutic agents.

“Pharmaceutically acceptable” refers to those compounds, materials,compositions, and/or dosage forms which are, within the scope of soundmedical judgment, suitable for contact with the tissues of human beingsand animals without excessive toxicity, irritation, allergic response,or other problem complications commensurate with a reasonablebenefit/risk ratio.

Examples of a pharmaceutically acceptable carrier include, for example,but not limited to, solvents, dispersion media, buffers, coatings,antibacterial and antifungal agents, wetting agents, preservatives,buggers, chelating agents, antioxidants, isotonic agents and absorptiondelaying agents.

Examples of a pharmaceutically acceptable carrier also include, forexample, but not limited to, water; saline; phosphate buffered saline;dextrose; glycerol; alcohols such as ethanol and isopropanol; phosphate,citrate and other organic acids; ascorbic acid; low molecular weight(less than about 10 residues) polypeptides; proteins, such as serumalbumin, gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides, andother carbohydrates including glucose, mannose, or dextrins; EDTA; saltforming counterions such as sodium; and/or nonionic surfactants such asTWEEN, polyethylene glycol (PEG), and PLURONICS; isotonic agents such assugars, polyalcohols such as mannitol and sorbitol, and sodium chloride;as well as combinations thereof.

Within the present invention, the disclosed compounds may be prepared inthe form of pharmaceutically acceptable salts. “Pharmaceuticallyacceptable salts” refer to derivatives of the disclosed compoundswherein the parent compound is modified by making acid or base saltsthereof. Examples of pharmaceutically acceptable salts include, but arenot limited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts or the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, such conventional non-toxic salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,ethane disulfonic, oxalic, isethionic, and the like. Thesephysiologically acceptable salts are prepared by methods known in theart, e.g., by dissolving the free amine bases with an excess of the acidin aqueous alcohol, or neutralizing a free carboxylic acid with analkali metal base such as a hydroxide, or with an amine.

Compounds described herein can be prepared in alternate forms. Forexample, many amino-containing compounds can be used or prepared as anacid addition salt. Often such salts improve isolation and handlingproperties of the compound. For example, depending on the reagents,reaction conditions and the like, compounds as described herein can beused or prepared, for example, as their hydrochloride or tosylate salts.Isomorphic crystalline forms, all chiral and racemic forms, N-oxide,hydrates, solvates, and acid salt hydrates, are also contemplated to bewithin the scope of the present invention.

Certain acidic or basic compounds of the present invention may exist aszwitterions. All forms of the compounds, including free acid, free baseand zwitterions, are contemplated to be within the scope of the presentinvention. It is well known in the art that compounds containing bothamino and carboxy groups often exist in equilibrium with theirzwitterionic forms. Thus, any of the compounds described herein thatcontain, for example, both amino and carboxy groups, also includereference to their corresponding zwitterions.

The term “stereoisomers” refers to compounds that have identicalchemical constitution, but differ as regards the arrangement of theatoms or groups in space. The term “enantiomers” refers to stereoisomersthat are mirror images of each other that are non-superimposable.

The term “administering” means either directly administering a compoundor composition of the present invention, or administering a prodrug,derivative or analog which will form an equivalent amount of the activecompound or substance within the body.

The term “inhibitor” as used herein includes compounds that inhibit theexpression or activity of a protein, polypeptide or enzyme and does notnecessarily mean complete inhibition of expression and/or activity.Rather, the inhibition includes inhibition of the expression and/oractivity of a protein, polypeptide or enzyme to an extent, and for atime, sufficient to produce the desired effect.

While not intending to be bound by any particular mechanism ofoperation, it is believed that the tyrosine hydroxylase inhibitorsaccording to the present invention function by decreasing the amount ofadrenaline secreted into the bloodstream.

The tyrosine hydroxylase inhibitor is well known in the art and fullydescribed in, for example, U.S. Patent Application Publications US2015/0290279, US 2015/0216827, US 2015/0111937, US 2015/0111878, US2013/0184214, and US 20130183263; U.S. Pat. Nos. 8,481,498, 9,308,188,and 9,326,962; and PCT Patent Application Publication WO2015061328,which are incorporated by reference herein in their entirety. Anysuitable tyrosine hydroxylase inhibitor, known to one of skilled in theart, can be used.

In certain embodiments, the tyrosine hydroxylase inhibitor is a tyrosinederivative. The tyrosine derivative can be capable of existing indifferent isomeric forms, including stereoisomers and enantiomers. Thetyrosine derivative can, for example, exist in both L-form or D-form.The tyrosine derivative can, for example, also exist in a racemic form.

Representative tyrosine derivatives include, for example, one or more ofmethyl (2R)-2-amino-3-(2-chloro-4 hydroxyphenyl) propanoate, D-tyrosineethyl ester hydrochloride, methyl(2R)-2-amino-3-(2,6-dichloro-3,4-dimethoxyphenyl) propanoateH-D-tyrosine(tBu)-allyl ester hydrochloride, methyl(2R)-2-amino-3-(3-chloro-4,5-dimethoxyphenyl) propanoate, methyl(2R)-2-amino-3-(2-chloro-3-hydroxy-4-methoxyphenyl) propanoate, methyl(2R)-2-amino-3-(4-[(2-chloro-6-fluorophenyl) methoxy] phenyl)propanoate, methyl (2R)-2-amino-3-(2-chloro-3,4-dimethoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-5-fluoro-4-hydroxyphenyl)propanoate, diethyl 2-(acetylamino)-2-(4-[(2-chloro-6-fluorobenzyl) oxy]benzyl malonate, methyl (2R)-2-amino-3-(3-chloro-4-methoxyphenyl)propanoate, methyl (2R)-2-amino-3-(3-chloro-4-hydroxy-5-methoxyphenyl)propanoate, methyl(2R)-2-amino-3-(2,6-dichloro-3-hydroxy-4-methoxyphenyl) propanoate,methyl (2R)-2-amino-3-(3-chloro-4-hydroxyphenyl) propanoate,H-DL-tyrosine methyl ester hydrochloride, H-3,5-diiodo-tyrosine methylester hydrochloride, H-D-3,5-diiodo-tyrosine methyl ester hydrochloride,H-D-tyrosine methyl ester hydrochloride, D-tyrosine methyl esterhydrochloride, D-tyrosine-methyl ester hydrochloride, methylD-tyrosinate hydrochloride, H-D-tyrosine methyl ester hydrochloride,D-tyrosine methyl ester hydrochloride, H-D-tyrosine methylester-hydrochloride, (2R)-2-amino-3-(4-hydroxyphenyl) propionic acid,(2R)-2-amino-3-(4-hydroxyphenyl) methyl ester hydrochloride, methyl(2R)-2-amino-3-(4-hydroxyphenyl) propanoate hydrochloride, methyl(2R)-2-azanyl-3-(4-hydroxyphenyl) propanoate hydrochloride,3-chloro-L-tyrosine, 3-nitro-L-tyrosine, 3-nitro-L-tyrosine ethyl esterhydrochloride, DL-m-tyrosine, DL-o-tyrosine, Boc-tyrosine (3,542)-OSu,Fmoc-tyrosine(3-NO2)-OH, α-methyl-L-tyrosine, α-methyl-D-tyrosine, andα-methyl-DL-tyrosine. In certain embodiments of the invention, thetyrosine derivative is α-methyl-L-tyrosine as shown below:

In other embodiments, the tyrosine derivative is α-methyl-D-tyrosine. Inother embodiments, the tyrosine derivative is α-methyl-DL-tyrosine in aracemic form as shown below:

In a particular embodiment, the tyrosine derivative is a structuralvariant of α-methyl-L-tyrosine or α-methyl-DL-tyrosine. The structuralvariants of α-methyl-L-tyrosine or α-methyl-DL-tyrosine are well knownin the art and fully described in, for example, U.S. Pat. No. 4,160,835,which is incorporated by reference herein in its entirety.

In one embodiment, the tyrosine derivative of the invention is anarylalanine compound having the formula:

-   -   wherein R₁ is hydrogen, methyl or ethyl ester group, or alkyl of        from 1 to 4 carbon atoms; R₂ is hydrogen, lower alkyl, lower        alkene, succinimide, or alkyl of from 1 to 4 carbon atoms; R₃ is        a substituted benzene ring of the following general formula

-   -   wherein Y₁, is located at the para position and is hydrogen,        hydroxy, methyl ether, dimethyl ether, trimethyl ether, or an        unsubstituted or halogen-substituted benzyl; Y₂, and Y₃ are the        same or different and wherein one or both Y₂, and Y₃ located at        either meta position or ortho position, and wherein Y₂, and Y₃        are hydrogen, hydroxy, halogen, methyl ether, or nitro; and R₄        is hydrogen, acetyl, tert-butyloxycarbonyl or        fluorenylmethyloxycarbonyl.

In some embodiments, Y1 and Y2 are the same or different and areselected from hydrogen, cyanoamino, carboxyl, cyano, thiocarbamoyl,aminomethyl, guanidino, hydroxy, methanesulfonamido, nitro, amino,methanesulfonyloxy, carboxymethoxy, formyl, methoxy and a substituted orunsubstituted 5- or 6-membered heterocyclic ring containing carbon andone or more nitrogen, sulfur or oxygen atoms, specific examples of suchheterocyclic rings being pyrrol-1-yl, 2-carboxypyrrol-1-yl,imidazol-2-ylamino, indol-1-yl, carbazol-9-yl,4,5-dihydro-4-hydroxy-4-trifluoromethylthiazol-3-yl,4-trifluoromethylthiazol-2-yl, imidazol-2-yl and4,5-dihydroimidazol-2-yl, such that (a) Y1 and Y2 cannot both behydroxy, (b) Y1 and Y2 cannot both be hydrogen and (c) when one of Y1and Y2 is hydrogen, the other cannot be hydroxyl.

In one example, R₃ is a substituted or unsubstituted benzoheterocyclicring having the formula:

-   -   in which the benzoheterocyclic ring is selected from the group        consisting of indolin-5-yl,        1-(N-benzoylcarbamimidoyl)-indolin-5-yl,        1-carbamimidoylindolin-5-yl, 1H-2-oxindol-5-yl, indol-5-yl,        2-mercaptobenzimidazol-5(6)-yl, 2-aminobenzimidazol-5(6)-yl,        2-methanesulfonamido-benzimidazol-5(6)-yl,        1H-benzoxazol-2-on-6-yl, 2-aminobenzothiazol-6-yl,        2-amino-4-mercaptobenzothiazol-6-yl, 2,1,3-benzothiadiazol-5-yl,        1,3-dihydro-2,2-dioxo-2,1,3-benzothiadiazol-5-yl,        1,3-dihydro-1,3-dimethyl-2,2-dioxo-2,1,3-benzothiadiazol-5-yl,        4-methyl-2(1H)-oxoquinolin-6-yl, quinoxalin-6-yl,        2-hydroxquinoxalin-6-yl, 2-hydroxyquinoxalin-7-yl,        2,3-dihydroxyquinoxalin-6-yl and        2,3-dihydro-3(4H)-oxo-1,4-benzoxazin-7-yl.

In another example, R₃ is a substituted or unsubstituted heterocyclicring having the formula:

-   -   in which the heterocyclic ring is selected from the group        consisting of 5-hydroxy-4H-pyran-4-on-2-yl, 2-hydroxypyrid-4-yl,        2-aminopyrid-4-yl, 2-carboxypyrid-4-yl, or        tetrazolo[1,5-a]pyrid-7-yl.

In one particular embodiment, the tyrosine hydroxylase inhibitor isaquayamycin. In one example, aquayamycin is a compound of the formulaset forth below.

In another particular embodiment, the tyrosine hydroxylase inhibitor isoudenone. In one example, oudenone is a compound of the formula setforth below.

Other suitable tyrosine hydroxylase inhibitor, known to one of skilledin the art, can also be used. Example of other tyrosine hydroxylaseinhibitor include, for example, but not limited to, cycloheximide,anisomycin, 3-iodo-L-tyrosine, pyratrione, phenyl carbonyl derivativeshaving catechol or triphenolic ring systems, for example, phenethylamineand gallic acid derivatives, 4-isopropyltropolone,2-(4-thiazolyl)benzimidazole, 8-hydroxyquinoline, o-phenantroline,5-iodo-8-hydroxyquinoline, bilirubin, 2,9-dimethyl-1, 10-phenantroline,α-α′-dipyridil, dibenzo [f,h]quinoxaline, 2,4,6-tripyridil-s-triazine,ethyl 3-amino4H-pyrrolo-isoxazole-5(6H)-carboxylate,α-nitroso-β-naphthol, sodium diethyldithiocarbamate,ethylenediamineteraacetic acid (See R Hochster, Metabolic Inhibitors V4:A Comprehensive Treatise 52 Elsevier (2012)).

In a particular embodiment, the tyrosine hydroxylase inhibitor is L1:79,which refers to α-methyl-DL-tyrosine in a racemic form or D,Lα-methyl-para-tyrosine (abbreviated as DL-AMPT).

α-methyl-DL-tyrosine inhibits the activity of tyrosine hydroxylase (TH),which catalyzes the first transformation in catecholamine biosynthesis,i.e., the conversion of tyrosine to dihydroxyphenylalanine (DOPA), whichis the rate limiting step in catecholamine synthesis.α-methyl-DL-tyrosine is a tyrosine analog that competes competitivelyfor TH and is excreted mostly unchanged in the urine.

In preliminary testing, it was observed that Demser (i.e.,α-methyl-L-tyrosine) had a beneficial effect on the symptoms of leakygut, but that this effect was short lived. The racemic mixture (i.e.,α-methyl-DL-tyrosine) was observed to have a greater physiologic effectand half-life. This may be due to the failure of the D amino acid isomerto be transported by the different L amino acid transport systems thatexist within the body, or by completion for renal excretion that allowsthe L isomer to persist for a longer duration.

In another aspect, the invention provides a pharmaceutical compositioncomprising a therapeutically effective amount or dose of a tyrosinehydroxylase inhibitor (e.g., α-methyl-DL-tyrosine) and apharmaceutically acceptable carrier.

Effective doses of the compositions of the present invention, fortreatment of autism as described herein vary depending upon manydifferent factors, including means of administration, target site,physiological state of the patient, whether the patient is an adult or achild, other medications administered, and whether treatment isprophylactic or therapeutic. Usually, the patient is a human butnon-human mammals including transgenic mammals can also be treated.Treatment dosages may be titrated using routine methods known to thoseof skill in the art to optimize safety and efficacy.

The pharmaceutical compositions of the invention may include a“therapeutically effective amount.” A “therapeutically effective amount”refers to an amount effective, at dosages and for periods of timenecessary, to achieve the desired therapeutic result. A therapeuticallyeffective amount of a molecule may vary according to factors such as thedisease state, age, sex, and weight of the individual, and the abilityof the molecule to elicit a desired response in the individual. Atherapeutically effective amount is also one in which any toxic ordetrimental effects of the molecule are outweighed by thetherapeutically beneficial effects.

As used herein, the terms “treat” and “treatment” refer to therapeutictreatment, including prophylactic or preventative measures, wherein theobject is to prevent or slow down (lessen) an undesired physiologicalchange associated with a disease or condition. Beneficial or desiredclinical results include, but are not limited to, alleviation ofsymptoms, diminishment of the extent of a disease or condition,stabilization of a disease or condition (i.e., where the disease orcondition does not worsen), delay or slowing of the progression of adisease or condition, amelioration or palliation of the disease orcondition, and remission (whether partial or total) of the disease orcondition, whether detectable or undetectable. Those in need oftreatment include those already with the disease or condition as well asthose prone to having the disease or condition or those in which thedisease or condition is to be prevented.

Dosage regimens may be adjusted to provide the optimum desired response(e.g., a therapeutic or prophylactic response).

In one example, a single bolus may be administered. In another example,several divided doses may be administered over time. In yet anotherexample, a dose may be proportionally reduced or increased as indicatedby the exigencies of the therapeutic situation. Dosage unit form, asused herein, refers to physically discrete units suited as unitarydosages for treating mammalian subjects. Each unit may contain apredetermined quantity of active compound calculated to produce adesired therapeutic effect. In some embodiments, the dosage unit formsof the invention are dictated by and directly dependent on the uniquecharacteristics of the active compound and the particular therapeutic orprophylactic effect to be achieved.

The composition of the invention may be administered only once, or itmay be administered multiple times. For multiple dosages, thecomposition may be, for example, administered three times a day, twice aday, once a day, once every two days, twice a week, weekly, once everytwo weeks, or monthly.

It is to be noted that dosage values may vary with the type and severityof the condition to be alleviated. It is to be further understood thatfor any particular subject, specific dosage regimens should be adjustedover time according to the individual need and the professional judgmentof the person administering or supervising the administration of thecompositions, and that dosage ranges set forth herein are exemplary onlyand are not intended to limit the scope or practice of the claimedcomposition.

“Administration” to a subject is not limited to any particular deliverysystem and may include, without limitation, oral (for example, incapsules, suspensions or tablets), parenteral (including subcutaneous,intravenous, intramedullary, intraarticular, intramuscular, orintraperitoneal injection), topical, or transdermal. Administration to ahost may occur in a single dose or in repeat administrations, and in anyof a variety of physiologically acceptable salt forms, and/or with anacceptable pharmaceutical carrier and/or additive as part of apharmaceutical composition (described earlier). Once again,physiologically acceptable salt forms and standard pharmaceuticalformulation techniques are well known to persons skilled in the art(see, for example, Remington's Pharmaceutical Sciences, Mack PublishingCo.).

In one aspect, the dosage of tyrosine hydroxylase inhibitor may rangefrom about 1 mg to about 4 g. In a particular embodiment, the dosage oftyrosine hydroxylase inhibitor may range from about 10 mg to about 1500mg. In some suitable embodiments of the invention, the compositioncomprises a tyrosine hydroxylase inhibitor (i.e., α-methyl-DL-tyrosine)in an amount ranging from about 50 mg (w/w) to about 1500 mg (w/w); fromabout 50 mg (w/w) to about 500 mg (w/w); from about 75 mg (w/w) to about350 mg (w/w); from about 90 mg (w/w) to about 350 mg (w/w); from about100 mg (w/w) to about 300 mg (w/w); or from about 100 mg (w/w) to about200 mg (w/w). In one embodiment, the composition comprises a tyrosinehydroxylase inhibitor (i.e., α-methyl-DL-tyrosine) in an amount of about90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 500, 750, 1000, or 1500mg (w/w/). As used herein, a “composition” refers to any compositionthat contains a pharmaceutically effective amount of one or more activeingredients (e.g., a tyrosine hydroxylase inhibitor, another acnetreating agent, or a combination thereof).

In some embodiments, a plurality of compositions having differentdosages are administered concurrently or sequentially. For instance, inone embodiment, a first composition comprising α-methyl-DL-tyrosine inan amount of about 200 mg (w/w) and a second composition comprisingα-methyl-DL-tyrosine in an amount of about 100 mg (w/w) are administeredconcurrently. In another embodiment, a first composition comprisingα-methyl-DL-tyrosine in an amount of about 200 mg (w/w) and a secondcomposition comprising α-methyl-DL-tyrosine in an amount of about 100 mg(w/w) are administered sequentially.

In one aspect, the composition is administered for at least about 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks. In another aspect, thecomposition is administered for a duration of 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, or 12 week dosing period. In yet another aspect, the compositionis administered for a duration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or12 month dosing period.

In another aspect, the invention provides a method for obtaining aplasma concentration of a therapeutic tyrosine hydroxylase inhibitordrug (e.g., α-methyl-DL-tyrosine) for a long term for treating an autismin a subject in need thereof, the method comprising administering tosaid subject said therapeutic drug at a concentration ranging from about50 mg (w/w) to about 500 mg (w/w) three times a day, wherein said plasmaconcentration ranges from about 500 ng/ml to 5000 ng/ml, and whereinsaid term is at least 1 week. In one embodiment, to obtain a desiredplasma concentration, the drug is administered for at least 1 day, 2day, 3 day, 4 day, 5 day, 6 day, 1 week, 2 week, 3 week, or 4 week. In aparticular embodiment, to obtain a desired plasma concentration, thedrug is administered for a duration ranging from about 1 day to about 4weeks; from about 2 days to about 4 weeks; or from about 1 week to about4 weeks. In some embodiments, to obtain a desired plasma concentration,the drug is administered for a duration of therapeutic regimens inexcess of 6 months.

The administration of composition of the invention may result in aplasma concentration ranging from about 500 ng/ml to 5000 ng/ml; fromabout 800 ng/ml to 2500 ng/ml, or from about 1400 ng/ml to 1800 ng/ml.

The pharmaceutical compositions of the invention may be formulated in avariety of ways, including for example, solid, semi-solid, and liquiddosage forms, such as capsules, tablets, pills, powders, liquidsolutions (e.g., injectable and infusible solutions), dispersions orsuspensions, liposomes and suppositories. In some embodiments, thecompositions are in the form of injectable or infusible solutions. Thecomposition is in a form suitable for oral, topical, intravenous,intraarterial, intramuscular, subcutaneous, parenteral, transmucosal, ortransdermal administration.

In a particular embodiment, the composition is in the form of a capsule.In another particular embodiment, the composition is in the form of atablet.

Administration of the pharmaceutical composition can be through variousroutes, including orally, nasally, subcutaneously, intravenously,intramuscularly, transdermally, vaginally, rectally or in anycombination thereof. Transdermal administration can be effected using,for example, oleic acid, 1-methyl-2-pyrrolidone, dodecylnonaoxyethylene.

In one aspect, the invention provides administering to a subject atherapeutically effective amount of a first tyrosine hydroxylaseinhibitor, for example, α-methyl-DL-tyrosine in combination with atherapeutically effective amount of a second tyrosine hydroxylaseinhibitor, for example, α-methyl-L-tyrosine. In another aspect, theinvention provides administering to a subject a therapeuticallyeffective amount of one or more tyrosine hydroxylase inhibitors, forexample, α-methyl-DL-tyrosine and/or α-methyl-L-tyrosine in combinationwith a therapeutically effective amount of another agent useful in thetreatment of autism.

In some of these aspects, 7-aminobutyric acid (GABA) can be administeredwith the tyrosine hydroxylase inhibitor (e.g., α-methyl-DL-tyrosine).The GABA can be administered simultaneously with the tyrosinehydroxylase inhibitor. In other aspects, the GABA can be administeredseparately from the tyrosine hydroxylase inhibitor, e.g., at anothertime during the day. In some aspects the GABA is administered atbedtime. Typically, dosages of the GABA are from about 5 mg to about 30mg, for example, 5, 10, 15, 20, 25, or about 30 mg of GABA, with 15 mgof GABA being particularly preferred.

In other of these aspects, a p450 3A4 promoter is administered inaddition to the tyrosine hydroxylase inhibitor (e.g.,α-methyl-DL-tyrosine) and the optional GABA. Preferred p450 3A4promoters include 5,5-diphenylhydantoin, valproic acid, andcarbamazepine.

Those subjects on the autism spectrum, including those diagnosed withAsperger's Disorder (Asperger Syndrome) or Social CommunicationDisorder, who also have symptoms of ADHD and/or tics can be treatedusing methods of the disclosure. In these aspects, the subject can beadministered an effective amount of a tyrosine hydroxylase inhibitor andan effective amount of a beta adrenergic agonist (also referred to asbeta agonists). The tyrosine hydroxylase inhibitor can be any of thetyrosine hydroxylase inhibitors described herein, withα-methyl-DL-tyrosine being particularly preferred. Beta adrenergicagonists are known in the art and include, for example, albuterol,levalbuterol, fenoterol, formoterol, isoproterenol, metaproterenol,salmeterol, terbutaline, clenbuterol, isoetarine, pirbuterol,procaterol, ritodrine, epinephrine, and combinations thereof. Albuterolis a particularly preferred beta adrenergic agonist.

In certain of these aspects, the tyrosine hydroxylase inhibitor and thebeta adrenergic inhibitor are administered simultaneously. In otheraspects, the beta adrenergic inhibitor is administered separately fromthe α-methyl-DL-tyrosine, e.g., at another time during the day.

According to the disclosure, the described methods for treating adisease or disorder can be used in combination with treatment methodsthat are also known to be effective in treating the same disease ordisorder. For example, autism behaviors and symptoms can be treated withcompounds that affect autonomic neurotransmission (e.g. amphetamine,methylphenidate, and the like), psychotopic drugs (e.g., risperidone),neutotransmitter reuptake inhibitors (e.g., fluoxetine), compounds thatstimulate glutaminergic transmission (e.g., LY2140023), and/or compoundsthat affect cholinergic neurotransmission (e.g., galantamine). As such,the disclosure is also directed to methods of treating autism in apatient by administering an effective amount of a tyrosine hydroxylaseinhibitor (e.g., α-methyl-DL-tyrosine) and an effective amount of acompound that affects autonomic neurotransmission, a psychotopic drug, aneutotransmitter reuptake inhibitor, a compound that stimulatesglutaminergic transmission, and/or a compound that affects cholinergicneurotransmission. In one embodiment, the invention provides methods oftreating autism in a patient by administering a therapeuticallyeffective amount of a tyrosine hydroxylase inhibitor (e.g.,α-methyl-DL-tyrosine) and a therapeutically effective amount of acentral nervous system (CNS) agent.

The diseases or disorders treated by the composition of the inventioninclude, for example, autism or its associated disease or disorder.

The autonomic nervous system has been implicated in symptoms thatresemble those seen in autism. Autism spectrum disorder (ASD) has beenassociated with abnormal findings in autonomic related structuresincluding the insula and the amygdala. Autonomic related changes such asincreases in basal heart rate and diminished heart rate due topsychosocial challenges are seen in autism. The autism-autonomic linkageis exemplified by the consequences of respiratory sinus arrhythmia (RSA)that includes difficulties with socialization, language difficulties,and delays in cognitive development.

It has been hypothesized a chronically over activated autonomic systemis a correlate of autism based upon the exaggerated levels of anxietythat attend autism, physiologic hyperarousal, and other correlates.Anxiety, perhaps the greatest co-morbidity associated with autism andwhich may drive other features of the disease, has been associated withcentral nervous system structures that are linked to autonomic function.Phenotypically autism and anxiety both present with stereotypedrepetitive behaviors and limited interests, avoidance behaviors, andspeech problems.

The relationship between anxiety and reported autonomic symptoms ofelevated heart rate, perspiration, and other sequelae of the “fight orflight” reaction reveal a role for the peripheral nervous systemfunction in autism. However, this may be secondary to central autonomicactivation. Central functions may manifest as elevated emotionalresponsiveness and exaggerated threat perception or diminishedinhibition of fear responses, which are associated with the centralstructures mentioned above in which autonomic responsiveness andemotional responsiveness overlap.

There is a considerable body of evidence that associates autism withcholinergic function in the central nervous system, specifically withvarious α-subtype nicotinic receptors, notably in the cerebellum.However, as autonomic function is classically considered to be a balanceof cholinergic and catecholaminergic systems, perceived increases ordecreases in cholinergic function may be manifestations of change in thedynamic balance of these systems with catecholaminergic tone. It isbelieved that it may be possible to effect therapeutic change in ASDthrough manipulation of either acetylcholine based manipulations or thecounterbalancing of dopamine, norepinephrine, or epinephrine mediatedmechanisms by administration of α-methyl-DL-tyrosine.

The present invention also relates to the treatment of the core symptomsof ASD by administration of α-methyl-DL-tyrosine to subjects in needthereof.

While not intending to be bound by any particular mechanism ofoperation, it is believed that the tyrosine hydroxylase inhibitor (e.g.,α-methyl-DL-tyrosine) administered according to the present inventionmodulates the catecholaminergic pathways implicated in autism, morespecifically such pathways involved in the core symptoms of ASD,including the catecholaminergic functions in the CNS and in thegastrointestinal tract. Therefore, the known effects of catecholamineson the endocrine and neuroendocrine systems are regulated byadministration of the tyrosine hydroxylase inhibitor (e.g.,α-methyl-DL-tyrosine), including but not limited to decreasing theamount of adrenaline secreted into the bloodstream, which may lessen theintensity of irritability and agitation, and other core symptoms of ASD.

In one aspect, the invention provides a method for treating coresymptoms of Autism Spectrum Disorder (ASD) in a subject in need thereof,the method comprising administering a therapeutically effective amountof a racemic mixture of α-methyl-DL-tyrosine.

In another aspect, the invention provides a method for treating anautism associated clinical trait in a subject in need thereof, themethod comprising administering to said subject a composition comprisinga therapeutically effective amount of a tyrosine hydroxylase inhibitor(e.g., α-methyl-DL-tyrosine), thereby treating said autism associatedclinical trait in said subject.

Examples of an autism associated clinical trait include, for example,but not limited to, a lack of social communication, a lack of socialinteraction, a lack of social motivation, lethargy and socialwithdrawal, inappropriate speech, hyperactivity, stereotypic behavior,irritability and agitation, restrictive behavior, repetitive behavior,ritualistic behavior, sameness behavior, compulsive behavior,self-injurious behavior or a combination thereof.

In one embodiment, the clinical trait is assessed based upon a changefrom a baseline in one or more psychometric tests. Examples of apsychometric test include, for example, but not limited to, clinicalglobal impression (CGI) rating scale, Vineland adoptive behavior scale(VABS), autism diagnostic observation schedule (ADOS), socialresponsiveness scale (SRS), aberrant behavior checklist-community(ABC-C), repetitive behavior scale (RBS), Conners parent rating scale(CPRS), or a combination thereof. In a particular embodiment, theclinical trait meets the requirements of Diagnostic and StatisticalManual of Mental Disorders-V (DSM-V) criteria.

The terms “subject,” “individual,” and “patient” are usedinterchangeably herein, and refer to an animal, for example a human, towhom treatment, including prophylactic treatment, with thepharmaceutical composition according to the present invention, isprovided. The term “subject” as used herein refers to human andnon-human animals. The terms “non-human animals” and “non-human mammals”are used interchangeably herein and include all vertebrates, e.g.,mammals, such as non-human primates, (particularly higher primates),sheep, dog, rodent, (e.g. mouse or rat), guinea pig, goat, pig, cat,rabbits, cows, horses and non-mammals such as reptiles, amphibians,chickens, and turkeys.

In one embodiment, the subject is a human patient between 3 years of ageand 21 years of age; 5 years of age and 21 years of age; or 6 years ofage and 17 years of age. In another embodiment, the subject is an adulthuman patient.

All patents and literature references cited in the present specificationare hereby incorporated by reference in their entirety.

Also provided herein are kits comprising one or more molecules orcompositions described herein. The following examples are provided tosupplement the prior disclosure and to provide a better understanding ofthe subject matter described herein. These examples should not beconsidered to limit the described subject matter. It is understood thatthe examples and embodiments described herein are for illustrativepurposes only and that various modifications or changes in light thereofwill be apparent to persons skilled in the art and are to be includedwithin, and can be made without departing from, the true scope of theinvention.

EXAMPLES Example 1

Two-hundred patients were initially screened. Thirty subjects meetingthe study criteria consented. Nine (9) subjects had high blood glucoselevels (hyperglycemia) prior to consenting to the study.

A high blood glucose level (hyperglycemia) is defined as a fastingplasma blood glucose level of 126 mg/dl or greater on two separateoccasions. The average patient age was sixty-two years old and themedian patient age was sixty years old. Six of the patients were femaleand three of the patients were male. Five of the patients were fifty tosixty years old and four of the patients were over the age of sixty.

The patients in the study were administered a treatment regimen thatincluded a tyrosine hydroxylase inhibitor (i.e., α-methyl-DL tyrosine),a melanin promoter (i.e., melanotan II), a p450 3A4 promoter (i.e.,5,5-diphenylhydantoin), and a leucine aminopeptidase inhibitor (i.e.,N-[(2S,3R)-3-amino-2-hydroxy-4-phenylbutyryl]-L-leucine). Thesecompounds were administered on each of five days per week for a periodof six weeks, with one or two days off between weekly cycles. Bloodglucose level was monitored for all subjects biweekly. Blood glucoselevels were determined by daily blood glucose tests followed-up withlaboratory blood glucose tests every two weeks.

After approximately two to four weeks, all nine of the subjects hadnormal blood glucose levels defined as a fasting plasma blood glucoselevel of 125 mg/dl or lower on two separate occasions.

Overall, the above-noted treatment was well tolerated by the subjects,with no adverse events related to the treatment, and responses have beendocumented to the treatment 100%.

Example 2

Patients are screened and the extent to which they meet the DSM-Vcriteria for autism spectrum disorder is assessed. A subgroup of thosesatisfying the criteria are administered a treatment regimen thatincludes a tyrosine hydroxylase inhibitor (i.e., α-methyl-DL tyrosine)at dose of 50-100 mg three times daily. Another subgroup is administereda treatment regimen that further includes a p450 3A4 promoter (i.e.,5,5-diphenylhydantoin) in one daily dose of 30 mg.

Gamma-aminobutyric acid is optionally administered to both subgroups atbedtime at a dose of 15 mg to aid sleeping and to quiet ticks andrepetitive behaviors like teeth grinding. Vasopressin is administered asneeded to assist brain governance. Following each administration of thetreatment regimen, changes in the extent to which the subjects satisfythe DSM-5 criteria is again assessed.

Example 3

A Blinded, Randomized, Placebo Controlled Phase 2 Study for the Use ofAMPT in the Treatment of Autism

Study Design: Blinded, randomized, 2 arm, 8-week treatment periodfollowed by additional follow-up visits off treatment over the next 18weeks for a total of 26 weeks' study participation. Treatment armsconsist of LI-79 alone or placebo.

Sample Size: L1-79 N=30, placebo N=10

Study Population: Autistic patients over 12 years of age that meet theentry criteria and who are high performing in the opinion of theinvestigator.

Major Inclusion Criteria: Signed informed consent, normal clinicallaboratory values, DSM-5 compliant diagnosis of autism, Qualifying ADOSscore, sufficiently high functioning to complete the protocol, nopsychotropic drugs for at least 2 weeks, ABC-C score >12.

Major Exclusion Criteria: Fragile-X syndrome, epilepsy, use ofcomplimentary alternative medications, any co-morbidities, otherpsychiatric disorder, out of range lab values.

Experimental Treatment: LI-79 is a racemic form of the drug Demser®. Itwill be given daily×8 weeks, and then followed on weeks 10, 13 and 26.

Non-Experimental Treatment: Placebo.

Dosage and Administration: LI-79 or placebo, as randomized, will beadministered orally at a dose of 90 mg TID.

Evaluation Schedule: Patients will receive 8 weeks of the schedule abovewith weekly treatment evaluations for weeks 1-8 then post-treatmentfollow up visits at weeks 10, 13 & 26.

Safety Measures: Regularly scheduled complete history and physicalexamination, vital signs, CBC, differential, platelet counts, urineanalysis, serum enzymes including: total protein, albumin, glucose, BUN,creatinine, direct and total bilirubin, alkaline phosphatase,phosphorous, calcium, aspartate aminotransferase (“AST”), alanineaminotransferase (“ALT”), sodium, potassium, chloride, bicarbonate, T₄,TSH, and adverse events assessments. An independent DMC will oversee theconduct of this trial to assure patient safety.

Study Duration: A maximum of 50 weeks (12 weeks' enrollment, 38 weeks'treatment and follow-up).

Study Endpoints: The primary end point will be the assessment of theattending physician as reflected in the Clinical Global Impressions(CGI) scale based upon changes from baseline in various psychometrictests, including the Aberrant Behavior Checklist-Community (ABC-C),Conners Parent Rating Scale and the Autism Diagnostic ObservationSchedule (ADOS), as well as from their personal observations in theclinic, and from videographic information taken at regularly scheduledclinic visits (per this protocol) and provided by caregivers over thecourse of this study.

Example 4

α-methyl-DL tyrosine (AMPT) was administered to 3 patients. This groupof patients was qualified under the DSM-5 definition of autism andtreated. See Table 1.

TABLE 1 Patient Dosage Adverse number Age Sex AMPT Events 01-001 3 Male90 TID None 01-002 15 Male 90 TID None 01-003 11 Male 90 TID None

The Aberrant Behavior Checklist-Community (ABC-C), Autism DiagnosticObservation Schedule (ADOS), Conners Parent Rating Scale (CPRS), andGeneral Clinical Impressions scale were used to assess the disease.Videos were taken at each visit.

Data for two of these patients over the first 3 weeks of thisobservation are presented in the tables below. Conners Patent RatingScale Data is depicted in FIG. 1. These clinical improvements appear tobegin quickly and are durable. Continued improvement over weeks andmonths has been observed. No adverse events have been reported otherthan mild tiredness on the first day of dosing in two patients.

TABLE 2 PT LR Date of Assessment Test Dimension initial Day 7 Day 14 Day21 Day 28 ABC-C TOTAL 114 71 55 53 53 irritability 21 16 16 16 16lethargy 31 12 8 7 7 stereotypy 13 5 0 0 0 hyperactivity 34 31 24 23 23speech 5 7 7 7 7 DSM-V TOTAL 42 27 21 18 18 social 9 5 3 2 2communication 10 4 3 2 2 relationships 11 4 3 2 2 behavior 12 12 12 1212

TABLE 3 PT RS Date of Assessment Test Dimension initial Day 7 Day 14 Day21 Day 28 ABC TOTAL 53 28 20 20 20 irritability 3 3 1 1 1 lethargy 6 3 11 1 stereotypy 16 11 9 9 9 hyperactivity 20 8 6 6 6 speech 8 3 3 3 3 DSMV TOTAL 21 15 8 6.5 5.5 social 5 3 1.5 1 1 communication 5 4 1.5 1.5 1.5relationships 1 0 0 0 0 behavior 10 8 5 4 3

TABLE 4 PT WC Date of Assessment Test Dimension initial Day 7 Day 14 Day21 Day28 ABC-C TOTAL 70 30 18 17 17 irritability 16 4 2 2 2 lethargy 219 3 2 2 stereotypy 12 7 6 6 6 hyperactivity 20 10 7 7 7 speech 1 0 0 0 0DSM-V TOTAL 26 17 14 12 11 social 6 6 4 2 2 communication 3 0 0 0 0relationships 6 2 2 2 2 behavior 11 9 8 8 7

Example 5 Evaluation of L1-79 Administration in Patients with Autism

L1-79 was used anecdotally in two patients followed by a more structuredevaluation in 8 additional patients with autism. All patients wereadministered a starting dose of L1-79 of 90 mg TID for a minimum of 3months. During the evaluation doses as high as 400 mg TID were used. Asummary of data available on these 10 patients was provided in theSummary of Clinical Efficacy, Use of L1-79 to treat autism, submitted toIND 128673, sequence number 0005. While the sample size was small andinvolved open-label administration of L1-79, the results wereencouraging with consistent improvements in the ABC-C domains and anaverage reduction in the ADOS-2 of 30%, and one child manifesting anADOS-2 decrease of 60% which resulted in the loss of his autismdiagnosis. In addition, longer treatment resulted in a greater magnitudeof therapeutic benefits on the core symptoms of autism. Moreover, whenthe study drug, L1-79, was discontinued not all of the benefitsregressed. These data suggest that L1-79 has the potential to improvethe core symptoms of autism.

Example 6 A Randomized, Double-Blind Placebo-Controlled 4-week Study inMale Patients Diagnosed with Autism

The purpose of this clinical study (referred to herein as “Study HT02-121”; i.e., Example 6) was to determine whether L1-79 was awell-tolerated and clinically useful agent for the treatment of ASD, andto assess the PK and pharmacodynamics (PD) of four weeks of TID dosingwith L1-79. While the preliminary study of Example 5 involved open-labeltreatment with L1-79 for at least three months, the present study wasonly 4-weeks based upon limitations in the available toxicology data.Based on the shorter duration, the response to L1-79 was expected to beless in Study HT 02-121 compared to the preliminary study of longerduration.

This clinical study was a randomized double-blind, placebo-controlledtwo-cohort, 4-week dose-escalation study that incorporated twoopen-label treatment groups to assess the safety and efficacy of L1-79100 mg and 200 mg TID in male patients between the ages of 12-21 yearsof age diagnosed with autism. The first cohort of 20 patients wascomprised of three groups of patients: The first group of five patientsreceived open-label L1-79 100 mg TID and underwent PK and EKGassessments. The remaining two groups in this cohort consisted of 15patients randomized on a 2:1 basis to receive L1-79 100 mg TID orplacebo. The PK and safety data from the open-label L1-79 100 mg TIDgroup was submitted to FDA for review and acceptance before the secondcohort was enrolled. The second cohort was procedurally identical to thefirst. The second cohort of 20 patients was comprised of the same threegroups of patients but a dose of 200 mg TID was used instead of 100 mgTID. The key inclusion criteria were as follows: (1) males between 13and 21 years of age, (2) DSM-5 compliant diagnosis of autism spectrumdisorder, confirmed by the Autistic Diagnosis Interview Review (ADIR),and by an ADOS-2 score consistent with a diagnosis of autism, (3) musthave been stable on no more than one concomitant medication and noplanned changes in psychosocial interventions during the study.

The key exclusion criteria were as follows: (1) sexually active males,(2) a history of Fragile-X syndrome, Rett syndrome or any otherco-morbidity including but not limited to cancer, genetic diseases, orany disease or syndrome that required drug therapy, (3) DSM-5 diagnosisof schizophrenia, schizoaffective disorder, alcohol use disorder orattention deficit hyperactivity disorder (ADHD), (4) had any activemedical problem(s), including epilepsy and asthma, (5) uncontrolledintercurrent illness including, but not limited to, ongoing or activeinfection, symptomatic cardio-vascular disease, hepatic disease, renaldisease, skeleto-muscular disease, human immunodeficiency virus (HIV),hemorrhagic cerebrovascular accident (HCVA), hepatitis B virus (HBV), orpsychiatric illness/social situations that would limit compliance withstudy requirements, (6) any disease that required chronic treatment, (7)any disease that required treatment with an immunosuppressive drug,and/or (8) current or lifetime diagnosis of severe psychiatric disorder.

Open-Label Patients

The first five patients in each cohort were assigned to receive activemedication in an open-label fashion. The patients were treatedidentically to the randomized patients with the following exceptions:(1) blood samples were drawn at baseline and 1 hour after dosing, and atWeek 1, 2, 3 and 4 treatment visits 1 hour after dosing to determine thePK of L1-79, (2) EKGs were assessed at baseline and within 3-days priorto the Week 1, 2, 3 and 4 treatment visits as well as the 1-week and4-week post-dosing follow-up visits, (3) vital signs, physical exams,and ASD assessments were performed at 1-week postdosing visit, (4) bloodand urine samples were drawn at baseline and at Week 1, 2, 3 and 4treatment visits 1 hour after dosing as well as the 1-week and 4-weekpost-dosing follow-up visits for safety analyses.

Randomized Patients

The randomized patients were treated identically to the open-labelpatients with the following three exceptions: (1) no blood samples forPK were drawn, (2) no ECG was performed, and (3) these patients did nothave a 1-week follow-up visit, only a 4-week follow-up visit. A time andevents schedule for the study is provided below in Table 5.

TABLE 5 Time and Events Schedule Visit Wk 0 1 Wk Post 4 Wk Form # FormName Screening Baseline Wk 1 Wk 2 Wk 3 Wk 4 Dosing FUP Study Day 0 ± 2 7± 2 14 ± 2 21 ± 2 28 ± 2 35 ± 2 56 ± 2 1 Date of Visit x x x x x x x x 2Informed Consent & Study Compliance x 3 Demography x 4 AdditionalPatient Information x 5 Inclusion Criteria x 6 Exclusion Criteria x 7Eligibility Criteria x 8 Medical History x 9 Family History x 10 Lab -Hematology (All Patients) x x x 11 Lab - Serum Chemistry (All Patents xx x 12 Urine Analysis (All Patients) x x x 13 Lab - Hematology (AssignedPatients Only) x* x x x x x** x 14 Lab - Serum Chemistry (AssignedPatients Only) x* x x x x x** x 15 Urine Analysis (Assigned PatientsOnly) x* x x x x x** x 16 PK/PD x* x x x x 17 Blood Draw x x x** x 18Vital Signs - Screening x 19 Vital Signs x x x x x x** x 20 PhysicalExamination x x** x 21 Randomization x 22 ADOS x x 23 EKG (AssignedPatients Only) x x x x x x** x 24 ADI-R (Telephonic) x 25 SAS x x** x 26VABS II x x x** x 27 ABC-C |All Scales| x x x x x x** x 28 SRS x x x x xx** x 29 RBS-R x x x x x x** x 30 CGI-S x x x x x x** x 31 CGI-I x x x xx** x 32 CGI-Overall Score x x x x x x** x 33 Study Drug Dispensing x x34 Study Drug Administration x x x x 35 Drug Accountability x x 36Adverse Event x x x x x x** x 37 Prior and Concomitant Medication x x xx x x x** x *At baseline visit for Assigned Group Patients PK samplesare to be taken before, and 1 hour after, the 1^(st) dose. **AssignedGroup Patients OnlySafety Endpoints

Safety was the primary endpoint of the study. The following safetyendpoints were assessed: ECGs, physical exams, laboratory evaluations(hematology, chemistry and urinalysis), vital signs, includingorthostatic blood pressure, adverse events (AEs) and concomitantmedications.

Efficacy Endpoints

The primary efficacy endpoint was the determination of the clinicalimprovement by the investigator as documented by the Clinical GlobalImpression (CGI) rating scales. Additional efficacy endpoints included:(1) changes from baseline in the Socialization and Communication Domainsof the Vineland Adaptive Behavior Scales, Second Edition (VABS II)parent/caregiver rating form, (2) changes from baseline in the ADOS-2Total Score and subscores, (3) changes in the Social ResponsivenessScale, Second Edition (SRS-2) Total Score and subscales, (4) changes inthe ABC-C domains, (5) changes in the Repetitive Behavior Scale-Revised(RBS-R) Total Score and subscales. Given the exploratory design of thisstudy only descriptive statistics were planned.

Results

Study Population

Patient enrollment and disposition are summarized in FIG. 2. A total of42 patients were randomized and received at least one dose of studydrug. One patient was randomized to open-label L1-79 200 mg TID but theparent requested voluntary withdrawal from the study at Week 0/Baselineafter receiving a single dose of study medication in clinic. Thirty-ninepatients completed the study. One patient treated with double-blindL1-79 200 mg TID withdrew from the study due to an AE (see Table 8) andone patient treated with placebo voluntarily withdrew from the study.Demographic characteristics are summarized in Table 6.

TABLE 6 Demographic Characteristics L1-79 100 mg TID L1-79 200 mg TIDOpen label Double-blind Open label Double blind Placebo N = 5 N = 10² N= 5¹ N = 11 N = 10² Age (years) Mean (SD) 16.2 (2.9) 16.4 (2.2) 16.4(1.1) 16.4 (2.7) 15.8 (2.7) Range (min, max) 13, 19 13, 20 15, 18 12, 2012, 19 Sex, n (%) Male 5 (100) 10 (100) 5 (100) 11 (100) 10 (100) Race,n (%) Caucasian 4 (80) 10 (100) 4 (80) 9 (82) 7 (70) African American 00 1 (20) 0 2 (20) Other 1 (20) 0 0 2 (18) 1 (10) ¹One patient wasrandomized to open-label L1-79 200 mg TID and only received a singledose of study medication. Data from this patient are not included in thetable. ²There was apparent confusion in the assigned-vs.-received drugkit numbers for two consecutively enrolled patients at site 01 who wererandomized on the same day (subjects 01-008-01-009). Since there issufficient uncertainty about what treatment the two patients actuallyreceived, data was analyzed conservatively by omitting these twopatients from the efficacy analyses. Data from these two patients wereincluded in the study population and safety analyses (however, neithersubject had reported AEs.).SafetyConcomitant Medications and Physical Exams

The majority of patients were not on CNS medications during the study.The following CNS medications were used during the study: clonidine(n=2), Strattera® (n=1), Depakote® (n=3), lorazepam (n=1), Prozac®(n=1), and Abilify® (n=1). There were no clinical significant physicalexam findings reported during the study.

Adverse Events

The incidence of AEs by primary system organ class and preferred term ispresented in Table 7, and a listing of the AEs reported during the studyis presented in Table 8. All AEs were mild to moderate in intensity andself-limited.

TABLE 7 Incidence of Adverse Events by Primary System Organ Class andPreferred Term L1-79 100 mg TID L1-79 200 mg TID Open label Double-blindOpen label Double blind Placebo Primary SOC N = 5 N = 10 N = 6 N = 11 N= 10 Preferred Term n (%) [e] (%) n (%) [e] (%) n (%) [e] (%) n (%) [e](%) n (%) [e] (%) All primary SOC 2(40.0)[6](100) 1(10.0)[1](100)2(40.0)[4](100) 6(54.5)[10](100) 3(30.0)[6](100) GI disorders 0[0] 0[0]0[0] 3(27.3)[3](30.0) 0[0] Diarrhoea 0[0] 0[0] 0[0] 3(27.3)[3](30.0)0[0] General disorders 0[0] 0[0] 2(40.0)[2](50) 0[0] 0[0] &administration site conditions Fatigue 0[0] 0[0] 1(20.0)[1](25.0) 0[0]0[0] Pyrexia 0[0] 0[0] 1(20.0)[1](25.0) 0[0] 0[0] Immune system 0[0]0[0] 1(20.0)[1](25.0) 0[0] 0[0] disorders Seasonal allergy 0[0] 0[0]1(20.0)[1](25.0) 0[0] 0[0] Infections and 0[0] 0[0] 0[0]2(18.2)[2](20.0) 1(10.0)[1](16.7) Infestations Ear infection 0[0] 0[0]0[0] 1(9.1)[1](10.0) 0[0] Nasopharyngifs 0[0] 0[0] 0[0] 0[0]1(10.0)[1](16.7) Otitis externa 0[0] 0[0] 0[0] 1(9.1)[1](10.0) 0[0]Investigations 0[0] 0[0] 0[0] 3(27.3)[3](30.0) 0[0] Amylase ↑ 0[0] 0[0]0[0] 0[0] 1(10.0)[1](16.7) Blood TSH ↑ 0[0] 0[0] 0[0] 1(9.1)[1](10.0)0[0] Crystal urine 0[0] 0[0] 0[0] 2(18.2)[2](20.0) 0[0] present Nervoussystem 0[0] 1(10.0)[1](100) 0[0] 1(9.1)[1](10.0) 0[0] disordersGeneralised tonic- 0[0] 0[0] 0[0] 1(9.1)[1](10.0) 0[0] cloric seizureSyncope 0[0] 1(10.0)[1](100) 0[0] 0[0] 0[0] Psychiatric 2(40.0)[6](100)0[0] 0[0] 1(9.1)[1](10.0) 1(10.0)[2](33.3) disorders Agitation 0[0] 0[0]0[0] 0[0] 1(10.0)[1](16.7) Anxiety 1(20.0)[1](16.7) 0[0] 0[0]1(9.1)[1](10.0) 0[0] Confusional state 1(20.0)[1](16.7) 0[0] 0[0] 0[0]0[0] Intentional self- 2(40.0)[2](33.3) 0[0] 0[0] 0[0] 1(10.0)[1](16.7)injury Irritability 2(40.0)[2](33.3) 0[0] 0[0] 0[0] 0[0] Renal andurinary 0[0] 0[0] 0[0] 0[0] 1(10.0)[2](33.3) disorders Urinary 0[0] 0[0]0[0] 0[0] 1(10.0)[2](33.3) incontinence Respiratory, 0[0] 0[0]1(20.0)[1](25.0) 0[0] 0[0] thoracic & mediastinal disorders Cough 0[0]0[0] 1(20.0)[1](25.0) 0[0] 0[0]

TABLE 8 Listing of Adverse Events Patient AE Verbatim Term Treatment IDTreatment (Preferred Term) Severity Emergent Relationship OutcomeDiscontinued 02-009 DB 100 mg TID syncope (syncope) mild Y not relatedresolved N 01-014 DB 200 mg TID crystal urine present mild N¹ notrelated resolved N (urine cystine crystal present) 01-016 DB 200 mg TIDdiarrhoea (diarrhea) moderate Y not related resolved N 01-018 DB 200 mgTID crystal urine present mild N¹ not related resolved N (urine cystinecrystal present) 02-018 DB 200 mg TID anxiety (anxious mood) mild Ypossibly related resolved N 02-018 DB 200 mg TID otitis externa (otitisexterna) mild Y not related resolved N 02-018 DB 200 mg TID diarrhoea(diarrhea) moderate Y possibly related resolved N 02-018 DB 200 mg TIDblood thyroid stimulating hormone moderate N¹ possibly related resolvedN increased (elevated TSH] 02-023 DB 200 mg TID generalized tonic-clonicseizer moderate Y possibly related resolved Y (grand mal seizure) 02-024DB 200 mg TID ear infection (earl infection) mild N² not relatedresolving N 02-024 DB 200 mg TID diarrhoea (diarrhea) mild Y not relatedresolved N 01-017 DB Placebo TID amylase increased (elevated amylase)mild N² not related resolved N 02-017 DB Placebo TID urinaryincontinence (bladder incontinence) mild Y possibly related resolved N02-017 DB Placebo TID urinary incontinence (bladder incontinence) mild Ypossibly related resolved N 02-017 DB Placebo TID nasopharyngitis(common cold) mild Y not related resolved N 02-019 DB Placebo TIDintentional self-injury mild Y possibly related unknown N (selfinjurious behavior) 02-019 DB Placebo TID agitation (agitation) mild Ypossibly related unknown N 02-001 OL 100 mg TID irritability(irritability) mild Y possibly related resolved N 02-001 OL 100 mg TIDintentional self-injury mild Y possibly related resolved N (selfinjurious behavior) 02-001 OL 100 mg TID confusional state (mentalconfusion) mild Y possibly related resolved N 02-001 OL 100 mg TIDanxiety (anxious) mild Y possibly related resolved N 02-002 OL 100 mgTID irritability (irritability) mild Y probably related unknown N 02-002OL 100 mg TID intentional self-injury mild Y probably related resolvingN (violent episodes of self injurious behavior) 01-011 OL 200 mg TIDcough (cough) moderate Y not related resolved N 01-011 OL 200 mg TIDpyrexia (fever) moderate Y not related resolved N 01-016 OL 200 mg TIDseasonal allergy (seasonal allergy) moderate Y not related not resolvedN 01-016 OL 200 mg TID fatigue (tiredness) mild Y possibly relatedresolved N DB = double-blind; OL = open-label ¹AE reported at 4-weekfollow-up visit ²AE reported pre-treatment

Of the patients who experienced an AE (34.1%, 14 of 41 patients), themajority experienced AEs of mild (24.4%, 10 of 41 patients) to moderate(14.6%, 6 of 41 patients) intensity. Only three AEs were reported bymore than one patient. Intentional self-injury was reported as atreatment emergent AE by two patients in the open-label 100 mg group andone patient in the double-blind placebo group. Irritability was reportedas a treatment emergent AE by two patients in the open-label 100 mggroup. For one of the patients reporting intentional self-injury andirritability (02-002), these symptoms emerged within a few days ofwithdrawal from active treatment (i.e., after completion of Week 4). Forthe other patient (02-001), symptoms began while on active drugtreatment. Urine cystine crystal present was reported as a non-treatmentemergent AE at the 4-week follow-up visit by two patients in thedouble-blind 200 mg group.

There were no serious adverse events (SAEs) or deaths reporting duringthe study. A total of 2 patients were withdrawn from the study due toadverse events: 1 patient in the double-blind 200 mg group experienced atreatment emergent AE of grand mal seizure and 1 patient in thedouble-blind placebo group experienced treatment emergent AEs ofintentional self-injury and agitation.

Vital Signs

Orthostatic vital signs were taken at each visit as described in StudyHT 02-121. There were no changes observed that met criteria fororthostatic hypotension (i.e., drop in systolic blood pressure of 20 mmHg, drop in diastolic blood pressure of 10 mm hg, or increases in heartrate of 30 beats per minute). A small number of subjects demonstratedasymptomatic drop in systolic and/or diastolic blood pressures, alongwith increase in heart rate at Week 1 or Week 2 of treatment, but theseresolved by Week 4 and were mild in nature (e.g., a drop of 10 mm Hg insystolic blood pressure, 5 mm Hg in diastolic pressure and increase of15-24 beats per minute in heart rate). FIG. 3 provides an example of thelargest change from baseline that was noted, and demonstrates that thesechanges were transient, falling far short of orthostatic criteria.

ECGS and Laboratory Evaluations

No clinically significant changes from baseline EKGs were noted in anyof the patients. Urinalysis revealed no findings related to clinicalsymptoms. Of note was the observation that half (50%) of the patients onL1-79 developed crystalluria compared to 30% of placebo treatedpatients. Of those that developed crystalluria during the study,approximately half (50%) had concentrated urine (specificgravity >1.025). Most of the crystalluria consisted of calcium oxalatecrystals. There were no symptoms associated with the crystalluria.

One patient developed asymptomatic transient mild elevation of amylaseat Week 4 (146 U/L, upper limit of normal [ULN] is 125 U/L), whichresolved by the 4-week follow up visit.

There were no significant elevations in AST or ALT or any otherchemistry parameters noted in the safety population.

Assessment of hematology parameters revealed no significant deviationsin the safety population.

Pharmacokinetics

Pharmacokinetics were assessed in the open-label 100 mg and 200 mg TIDpatients. On Day 1, L-79 concentrations were assessed one-hour postdose. Subsequently, a random L1-79 plasma concentration was assessed atWeek 1, Week 2, Week 3 and Week 4. At 1-hour post-dose on Day 1 plasmaconcentrations with 100 mg and 200 mg TID ranged from 0 (<2.5) to 736ng/mL and 23 to 1680 ng/mL, respectively. A summary of individualpatient and combined overall mean L1-79 random plasma concentrations areshown in FIG. 4. As shown in FIG. 5 and FIG. 6, overall, random plasmaL1-79 concentrations were relatively stable over Week 1 to Week 4.

Efficacy

At the time this study was initiated, there was a lack of long-term,juvenile and reproductive toxicology data. Applicants embarked on ashort study of 28 days in duration, with a limitation in the number,gender, and age of the patients allowed into the study. As a result,this study was not designed or powered to demonstrate statisticalsignificance on any of the efficacy endpoints. Thus, as expected none ofthe outcomes measures achieved statistical significance. What wasanticipated, and what was observed, were positive trends in a variety ofinstruments consistent with an improvement in the core symptoms ofautism. In fact, there were multiple efficacy measures demonstratingsimilar indicators of improvement in the treatment of target core socialdomains affected by ASD. In addition, efficacy data are only displayedfor patients that received blinded treatment.

Outcome Measures

There were some differences in how questionnaires were administeredduring Cohort 1 and Cohort 2 of the study. In the preliminary study, allforms were completed at the study site in the presence of theinvestigator. In Cohort 1 of the present study, the VABS—II, SRS-2,ABC-C and RBS-R were filled out at home by the patient/patient'sfamilies prior to treatment visits and returned at treatment visits inorder to expedite the execution of the study. The assessment proceduresfor VABS II, SRS-2, ABC-C and RBS-R during Cohort 1 were not done at thetreatment visits and as a result, families had difficulties filling outthe forms properly. In order improve the quality and completeness of thequestionnaires during Cohort 2, all questionnaires (with the exceptionof the ADOS-2 [administered by external certified ADOS-2 testadministrator for both cohorts] and CGI [completed by investigator inboth cohorts]) were completed by the principal investigator with theassistance of the patient/patient's families at the pre-specifiedtreatment visits. As the Cohort 2 data was more robust, the discussionof results focus on the comparison of L1-79 200 mg to placebo, with theexception of the ADOS-2 and CGI, where both L1-79 100 mg and 200 mg arecompared to placebo.

After the conclusion of the study, it was discovered that the basalanchors for the VABS-II were not always appropriately established. Inaddition, only two subdomains (communication and socialization) werecompleted for the patients, making it impossible to obtain domain ortotal scores. However, the results for the VABS II communication andsocialization domains are presented below.

A tabular summary of the efficacy measures is presented below (Table 9).For those measures demonstrating positive trends in favor of L1-79 threesets of figures are presented for each efficacy measure. The figuresconsist of the following: a) a line graph showing the comparative changebetween the 3 treatment groups from screening/baseline to 4 weeks(followed by a 4-week post treatment timepoint in some cases), b) anindividual patient response plot, and c) a responder analysis plot. Forthose efficacy measures not showing a positive trend in favor of L1-79,only a line graph showing the comparative change between the 3 treatmentgroups from screening/baseline to week 4 (followed by a 4-week posttreatment timepoint in some cases) is presented.

Responder Definition:

In addition to analyzing the data by observing the magnitude of changein each efficacy endpoint for patients exposed to active drug comparedto placebo, it is important to define a ‘responder’ population. Ingeneral, responder analyses are intended to focus on the number ofpatients demonstrating any benefit, rather than the overall change inthe studied population. Typically, responders are defined as patientswhose target symptoms demonstrate a pre-specified improvement in theefficacy endpoint. This definition is consistent with accepted standardsfor judging mid and long term outcomes for experimental treatmentsstudied in nearly every disorder, including autism. However, it does nottake into account the need for a refined definition in studies of shortduration for disorders, like autism, that have oscillations in symptomseverity. In ASD, children often go through periods of mild to moderateimprovement or worsening in their behavioral, social and emotionalsymptom regulation related to a variety of identifiable and as yetunidentified factors (Boso, 2010; Jyonouchi, 2011). Thus, in a study ofshort duration, a ‘baseline’ measure may in fact be at the apex,midpoint or nadir of one of these oscillations. If they are at an apexor mid-point when the target symptoms under study are measured atbaseline, then a short duration study may only have the opportunity todemonstrate initial efficacy by preventing the target symptom fromreturning back to its oscillatory nadir. To account for this probableoccurrence among some study patients, the responder definition includespatients demonstrating either short term stability or improvement in thetarget symptoms. Thus, for each efficacy endpoint demonstrating apositive trend, responder analyses were conducted as defined herein.

A tabular summary of the change from screening/baseline for efficacymeasures is provided in Table 9.

TABLE 9 Summary Comparisons of Mean Changes from Screening/BaselineBetween L1-79 and Placebo for Efficacy Measures at Week 4 or LOCF.Double-blind L1-79 Double-blind L1-79 100 mg TID 200 mg TID Placebo N =9 N = 11 N = 9 CGI-S (N) 9 11 9 Mean Change from Baseline (SD) −0.3(0.5) −0.8 (0.9) −0.3 (0.5) Range (min, max) −1, 0 −2, 0 −1, 0 Quartiles(25^(th), median, 75^(th)) −1, −0, 0 −2, −1, 0 −1, 0, 0 Treatment effectversus placebo (95% CI) −0.0 (−0.5, 0.5) −0.5 (−1.2, 0.2) N/A P-valuefor treatment effect 1.00 0.16 N/A CGI-I (N) 9 11 8 Mean Change fromBaseline (SD) −0.6 (0.7) −0.5 (0.8) −0.1 (0.8) Range (min, max) −1, 1−2, 0 −1, 1 Quartiles (25th, median, 75th) −1, −1, 0 −1, 0, 0 −1, 0, 1Treatment effect versus placebo (95% CI) −0.4 (−1.2, 0.4) −0.3 (−1.1,0.5) N/A P-value for treatment effect 0.27 0.40 N/A VABS II StandardizedSocialization Score (N) 5 7 4 Mean Change from Baseline (SD) −4.2 (12.1)7.6 (11.9) 0.8 (1.5) Range (min, max) −14, 9 0, 29 0, 3 Quartiles(25^(th), median, 75^(th)) −13, −12, 9 0, 1, 20 0, 0, 2 Treatment effectversus placebo (95% CI) −5.0 (−19.9, 10.0) 6.8 (−4.2, 17.8) N/A P-valuefor treatment effect 0.41 0.18 N/A VABS II Standardized CommunicationScore 8 7 4 (N) Mean Change from Baseline (SD) 10.1 (16.6) 2.7 (4.3) 6.5(9.3) Range (min, max) −3, 46 0, 12 0, 20 Quartiles (25^(th), median,75^(th)) −1, 5, 16 0, 1, 3 1, 3, 13 Treatment effect versus placebo (95%CI) 3.6 (−16.5, 23.8) −3.8 (−12.9, 5.3) N/A P-value for treatment effect0.70 0.37 N/A ADOS-2 Total Score (N) 9 10 8 Mean Change from Screening(SD) −0.6 (1.7) −1.0 (2.2) 0.0 (2.1) Range (min, max) −4, 2 −6, 2 −3, 4Quartiles (25^(th), median, 75^(th)) −1, 0, 0 −1, −1, 0 −1, −1, 1Treatment effect versus placebo (95% CI) −0.6 (−2.5, 1.4) −1.0 (−3.2,1.2) N/A P-value for treatment effect 0.56 0.34 N/A ADOS-2 Restrictedand Repetitive Behavior 9 10 7 (N) Mean Change from Screening (SD) 0.3(1.6) −1.0 (2.1) −0.1 (2.5) Range (min, max) −1, 4 −6, 1 −3, 4 Quartiles(25^(th), median, 75^(th)) −1, 0, 1 −2, −1, 0 −2, −1, 2 Treatment effectversus placebo (95% CI) 0.5 (−1.7, 2.7) −0.9 (−3.2, 1.5) N/A P-value fortreatment effect 0.65 0.45 N/A ADOS-2 Social Affect (N) 9 10 7 MeanChange from Screening (SD) −0.9 (2.0) 0.0 (2.1) 0.1 (1.1) Range (min,max) −5, 1 −4, 3 −1, 2 Quartiles (25^(th), median, 75^(th)) −2, 0, 1 −1,1, 1 −1, 0, 1 Treatment effect versus placebo (95% CI) −1.0 (−2.9, 0.8)−0.1 (−2.0, 1.7) N/A P-value for treatment effect 0.24 0.87 N/A SRS-2Total T-score (N) 8 11 9 Mean Change from Baseline (SD) −0.4 (6.2) −7.7(10.9) −5.2 (9.6) Range (min, max) −11, 10 −30, 3 −22, 5 Quartiles(25^(th), median, 75^(th)) −3, −1, 3 −19, −3, 0 −12, −2, 1 Treatmenteffect versus placebo (95% CI) 4.8 (−3.7, 13.4) −2.5 (−12.3, 7.3) N/AP-vatue for treatment effect 0.24 0.60 N/A SRS-2 DSM-5 Social,Communication and 9 11 9 Interaction T-score (N) Mean Change fromBaseline (SD) 0.6 (5.4) −7.6 (10.3) −4.2 (10.4) Range (min, max) −9, 8−28, 3 −24, 8 Quartiles (25^(th), median, 75^(th)) −3, 2, 5 −18, −3, 0−12, −1, 1 Treatment effect versus placebo (95% CI) 4.8 (−3.5, 13.1)−3.4 (−13.2, 6.4) N/A P-value for treatment effect 0.24 0.47 N/A SRS-2DSM-5 Restricted and Repetitive 9 11 8 Behavior T-score (N) Mean Changefrom Baseline (SD) −0.4 (9.3) −7.6 (13.2) −7.6 (10.8) Range (min, max)−17, 14 −34, 6 −30, 3 Quartiles (25^(th), median, 75^(th)) −3, 0, 5 −22,−3, 3 −12, −7, 1 Treatment effect versus placebo (95% CI) 7.2 (−3.2,17.6) −0.0 (−12.1, 12.1) N/A P-value for treatment effect 0.087 1.00 N/ASRS-2 Social Communication T-score (N) 9 11 9 Mean Change from Baseline(SD) 1.1 (7.1) −8.3 (11.0) −4.9 (10.6) Range (min, max) −9, 13 −30, 1−22, 6 Quartiles (25^(th), median, 75^(th)) −5, 0, 6 −16, −4, −1 −13, 0,3 Treatment effect versus placebo (95% CI) 6.0 (−3.0, 15) −3.4 (−13.6,6.9) N/A P-value for treatment effect 0.18 0.50 N/A SRS-2Social-Motivation T-score (N) 9 11 9 Mean Change from Baseline (SD) −1.4(9.2) −10.6 (12.6) −1.8 (9.9) Range (min, max) −15, 12 −30, 6 −16, 15Quartiles (25^(th), median, 75^(th)) −7, −4, 8 −21, −6, 0 −8, −2, 4Treatment effect versus placebo (95% CI) 0.3 (−9.2, 9.9) −8.9 (−19.7,2.0) N/A P-value for treatment effect 0.94 0.10 N/A ABC-C Lethargy andSocial Withdrawal (N) 9 11 9 Mean Change from Baseline (SD) −1.6 (6.7)−4.5 (6.7) −5.4 (6.4) Range (min, max) −15, 7 −19, 5 −14, 3 Quartiles(25^(th), median, 75^(th)) −2, −1, 1 −9, −2, 0 −11, −3, −1 Treatmenteffect versus placebo (95% CI) 3.9 (−2.6, 10.4) 1.0 (−5.2, 7.2) N/AP-value for treatment effect 0.23 0.74 N/A ABC-C Inappropriate Speech(N) 9 11 9 Mean Change from Baseline (SD) 0.0 (1.7) −2.0 (2.4) −1.6(2.7) Range (min, max) −2, 2 −7, 0 −8, 1 Quartiles (25^(th), median,75^(th)) −2, 0, 1 −4, −1, 0 −2, 0, 0 Treatment effect versus placebo(95% CI) 1.6 (−0.7, 3.8) −0.4 (−2.9, 2.0) N/A P-value for treatmenteffect 0.16 0.71 N/A ABC-C Hyperactivity and Noncompliance 9 11 8 MeanChange from Baseline (SD) 0.4 (3.9) −4.0 (9.1) −4.5 (6.0) Range (min,max) −8, 5 −28, 7 −13, 6 Quartiles (25^(th), median, 75^(th)) 0, 0, 3−7, −1, 0 −9, −5, −1 Treatment effect versus placebo (95% CI) 4.9 (−0.2,10.1) 0.5 (−7.3, 8.3) N/A P-value for treatment effect 0.060 0.89 N/AABC-C Irritability and Agitation 9 11 8 Mean Change from Baseline (SD)−0.9 (2.0) −2.2 (6.9) −3.4 (7.1) Range (min, max) −5, 2 −16, 9 −11, 12Quartiles (25^(th), median, 75^(th)) −2, −1, 0 −7 −2, 0 −8, −5, −1Treatment effect versus placebo (95% CI) 2.5 (−3.5, 8.5) 1.2 (−5.7, 8.0)N/A P-value for treatment effect 0.37 0.72 N/A ABC-C StereotypicBehavior 9 11 8 Mean Change from Baseline (SD) 1.9 (3.3) −1.7 (2.9) −0.6(5.4) Range (min, max) −1, 8 −7, 1 −5, 12 Quartiles (25^(th), median,75^(th)) 0, 0, 4 −3, −1, 0 −4, −2, −1 Treatment effect versus placebo(95% CI) 2.5 (−2.0, 7.1) −1.1 (−5.1, 2.9) N/A P-vatue for treatmenteffect 0.26 0.57 N/A RBS-R Total Score (N) 9 11 8 Mean Change fromBaseline (SD) −3.2 (7.8) −16.1 (24.1) −11.0 (11.0) Range (min, max) −14,12 −78, 1 −24, 6 Quartiles (25^(th), median, 75^(th)) −6, −4, −1 −28,−2, 0 −21, −12, −2 Treatment effect versus placebo (95% CI) 7.8 (−2.0,17.5) −5.1 (−22.7, 12.5) N/A P-value for treatment effect 0.11 0.55 N/ARBS-R Restricted Behavior (N) 9 11 7 Mean Change from Baseline (SD) 1.3(2.6) −2.5 (2.8) −1.6 (1.6) Range (min, max) −1, 8 −8, 0 −4, 0 Quartiles(25^(th), median, 75^(th)) 0, 1, 1 −5, −1, 0 −3, −2, 0 Treatment effectversus placebo (95% CI) 2.9 (0.5, 5.4) −1.0 (−3.4, 1.5) N/A P-value fortreatment effect 0.023 0.41 N/A RBS-R Ritualistic Behavior (N) 9 11 7Mean Change from Baseline (SD) 0.1 (2.8) −3.1 (4.1) −3.7 (3.5) Range(min, max) −4, 4 −13, 0 −9.0 Quartiles (25^(th), median, 75^(th)) −1, 0,2 −5, −1, 0 −7, −3, 0 Treatment effect versus placebo (95% CI) 3.8 (0.5,7.2) 0.6 (−3.4, 4.6) N/A P-value for treatment effect 0.028 0.74 N/ARBS-R Sameness Behavior (N) 9 11 7 Mean Change from Baseline (SD) −0.7(2.0) −4.2 (7.1) −2.9 (2.2) Range (min, max) −4, 2 −22, 1 −6, 0Quartiles (25^(th), median, 75^(th)) −1, −1, 0 −9, 0, 0 −5, −2, −1Treatment effect versus placebo (95% CI) 2.2 (−0.1, 4.4) −1.3 (−6.3,3.6) N/A P-value for treatment effect 0.056 0.57 N/A RBS-R CompulsiveBehavior (N) 9 11 7 Mean Change from Baseline (SD) −1.2 (3.6) −2.7 (4.9)−1.6 (1.7) Range (min, max) −8, 3 −16, 0 −4, 0 Quartiles (25^(th),median, 75^(th)) −2, 0, 1 −3, 0, 0 −3, −1, 0 Treatment effect versusplacebo (95% CI) 0.3 (−2.8, 3.5) −1.2 (−4.6, 2.3) N/A P-value fortreatment effect 0.82 0.49 N/A RBS-R Stereotypic Behavior (N) 9 11 7Mean Change from Baseline (SD) −0.6 (1.7) −1.8 (2.8) −0.4 (4.2) Range(min, max) −5, 1 −7, 0 −7, 6 Quartiles (25^(th), median, 75^(th)) 0, 0,0 −5, 0, 0 −3, 0, 3 Treatment effect versus placebo (95% CI) −0.1 (−4.1,3.8) −1.4 (−4.8, 2.1) N/A P-value for treatment effect 0.94 0.41 N/ARBS-R Self Injurious Behavior (N) 9 11 7 Mean Change from Baseline (SD)−2.2 (5.3) −1.7 (3.9) −2.1 (3.8) Range (min, max) −16, 1 −12, 1 −8, 2Quartiles (25^(th), median, 75^(th)) −2, 0, 0 −1, 0, 0 −7, 0, 0Treatment effect versus placebo (95% CI) −0.1 (−5.2, 5.0) 0.4 (−3.5,4.4) N/A P-value for treatment effect 0.97 0.83 N/A LOCF = lastobservation carried forwardClinical Global Impression (CGI) Rating Scales

The CGI-Overall Severity (CGI-S) and CGI-Overall Improvement (CGI-I) forL1-79 compared to placebo over time are displayed graphically in FIG. 7and FIG. 8. The mean CGI-S change from baseline at Week 4 or lastobservation carried forward (LOCF) is displayed in FIG. 9, anddemonstrates a mean 0.5 point improvement on the overall CGIS forpatients in the 200 mg group, compared to those in the placebo and 100mg groups.

A summary of the change in CGI-S from Baseline to Week 4 is displayed bypatient in FIG. 10. In addition, responder analyses (defined asimprovement) are presented for CGIS at Week 4 or LOCF in FIG. 11.

Responder definitions for CGI-S were defined as improvement only(instead of improvement or no change) since, unlike the other measureswhich were performed weekly, the CGI-S requires the clinician to give anoverall assessment of the patients' clinical symptom severity based onall of the outcome measures and their movement over the entirety of thestudy.

The responder analysis for CGI-S at Week 4 demonstrates a clear doseresponse trend of improvement for the L1-79 100 mg and 200 mg groupscompared to placebo.

Vineland Adaptive Behavior Scales, Second Edition (VABS II)

The change from Baseline in VABS II Standardized Socialization Score andthe VABS II Standardized Communication Score for L1-79 compared toplacebo over time are displayed graphically in FIG. 12 and FIG. 13,respectively.

Autism Diagnostic Observation Schedule, Second Edition (ADOS-2)

The change from Screening in ADOS-2 Total Score, Restrictive andRepetitive Behavior Total Score and Social Affect Total Score for L1-79compared to placebo over time are displayed graphically in FIG. 14, FIG.15 and FIG. 16. While not typically used as an outcome measure, thechange in ADOS-2 over a short period of time is consistent withopen-label preliminary study of longer duration previously presented.

The consistency of this effect across multiple patients is suggested bythe summary of the change in ADOS-2 Total Score from Screening to Week 4displayed in FIG. 17.

Social Responsiveness Scale, Second Edition (SRS-2)

The change from Baseline in SRS-2 Total T-score, SRS-2 DSM-5 Social,Communication and Interaction T-score, SRS-2 Social CommunicationT-score, SRS-2 Social-Motivation T-score and SRS-2 DSM-5 Restrictive andRepetitive Behavior T-score for L1-79 compared to placebo over time aredisplayed graphically in FIG. 18, FIG. 19, FIG. 20, FIG. 21 and FIG. 22,respectively.

The SRS-2 Total, SRS-2 DSM-5 Social Communication and Interaction, SRS-2Social Communication and SRS-2 Social Motivation T-scores improved byclose to 8 points or more on average in the L1-79 200 mg treated group(FIG. 18, FIG. 19, FIG. 20 and FIG. 21). The Social Communication andInteraction scale are comprised of the DSM-5 criteria that make up thesocial communication and social interaction deficits required for thediagnosis of ASD. Of considerable interest is the finding that patientswith baseline scores between 60 and 83 who dropped by 8 points or morewere likely to demonstrate categorical changes in clinical severity,since the classifications of “Within Normal Limits” (below 60 T-score),“Mild Range” (60 to 65 T-score), “Moderate Range” (66 to 75 T-score) and“Severe Range” (76 T-score or greater) of symptom severity are definedwithin those T-score ranges.

A summary of the change in SRS-2 Total T-score, SRS-2 DSM-5 Social,Communication and Interaction T-score, SRS-2 Social CommunicationT-score, and SRS-2 Social-Motivation T-score from baseline to Week 4 foreach patient are displayed in FIG. 23, FIG. 24, FIG. 25, and FIG. 26,respectively.

For SRS-2 Total, SRS-2 DSM-5 Social Communication and Interaction, SRS-2Social Communication, and SRS-2 Social Motivation T-scores, both themagnitude and number of responders observed were greater with L1-79 200mg, compared to placebo (FIG. 27, FIG. 28, FIG. 29 and FIG. 30). For theSRS-2 Total T-Score, 3 patients had sufficient improvement to changecategories of severity in the L1-79 200 mg group, compared to 2 in boththe L1-79 100 mg and placebo groups. For the SRS-2 Social MotivationT-score, 6 patients demonstrated categorical changes in clinicalseverity in the L1-79 200 mg group, compared to 4 and 3 patients in the100 mg and placebo groups, respectively. For the SRS-2 DSM-5 Social,Communication and Interaction T-score, 4 patients had changes inseverity categorization in the 200 mg group, compared to 2 and 3patients in the 100 mg and placebo groups, respectively. Similarly, forthe SRS-2 Social Communication T-score, 4 patients demonstratedcategorical improvement in the 200 mg group, compared to 3 in both theplacebo and 100 mg groups.

Responder analyses (defined as an improvement or no worsening) arepresented for SRS-2 Total T-score, SRS-2 DSM-5 Social, Communication andInteraction T-score, SRS-2 Social Communication T-score, and SRS-2Social-Motivation T-score at Week 4 or LOCF in FIG. 27, FIG. 28, FIG.29, and FIG. 30, respectively.

FIGS. 27-30 demonstrate that the percent of responders (as defined byimprovement or no worsening were greater with L1-79 200 mg, compared toplacebo; see Responder Definition (section above).

Aberrant Behavior Checklist-Community (ABC-C)

The change from Baseline in ABC-C Lethargy and Social Withdrawal Domain,ABC-C Inappropriate Speech Domain, ABC-C Hyperactivity and NoncomplianceDomain, ABC-C Stereotypic Behavior Domain and ABC-C Irritability andAgitation Domain for L1-79 compared to placebo over time are displayedgraphically in FIG. 31, FIG. 32, FIG. 33, FIG. 34 and FIG. 35,respectively.

A summary of the change in ABC-C Lethargy and Social Withdrawal Domainand ABC-C Inappropriate Speech Domain from Baseline to Week 4 aredisplayed by patient in FIG. 36 and FIG. 37, respectively. While therewas no separation between the 200 mg and placebo groups in mean changefrom baseline scores, responder analyses did demonstrate a greatertendency for patients to respond with improvement or no worsening ofbehaviors in the 200 mg group compared to placebo, as shown in FIG. 38and FIG. 39, respectively.

As with the SRS-2, both the number of responders and magnitude ofresponse for those patients given L1-79 200 mg demonstrated a trendtoward separation for the social and speech domains of the ABC-C.Equally notable was the observation that 90 to 100% of patients withL1-79 200 mg demonstrated stability or improvement of these domains,which are known to vary significantly in intensity over shorter periodsof time.

Repetitive Behavior Scale-Revised (RBS-R)

The change from Baseline in RBS-R Total Score, RBS-R RestrictiveBehavior, RBS-R Ritualistic Behavior, RBS-R Sameness Behavior, RBS-RCompulsive Behavior, RBS-R Stereotypic Behavior, and RBS-RSelf-injurious Behavior for L1-79 compared to placebo over time isdisplayed graphically in FIG. 40, FIG. 41, FIG. 42, FIG. 43, FIG. 44,FIG. 45, and FIG. 46.

There were mean improvements in RBS-R Total Score as noted by a decreasein score for L1-79 200 mg compared to placebo. (FIG. 40).

A summary of the change in RBS-R Total Score from Baseline to Week 4 isdisplayed by patient in FIG. 47. In addition, responder analyses(defined as an improvement or no worsening) are presented for RBS-RTotal Score at Week 4 or LOCF in FIG. 48.

Consistent with the ABC-C Lethargy and Social Withdrawal and ABC-CInappropriate Speech Domains, the RBS-R Total Score demonstrated a cleartrend toward a greater magnitude of reduction in restricted andrepetitive behaviors with L1-79 200 mg compared to placebo. This isfurther demonstrated by the responder analysis, which shows that 91% ofpatients treated with L1-79 200 mg had improvement or stabilization ofsymptoms compared to 78% with placebo.

Overall Conclusions

Currently available therapies for children and adults with ASD onlytarget collateral symptoms associated with the disorder (irritability,agitation, impulsivity, hyperactivity) and have side effects thatrequire monitoring metabolic parameters through blood tests. Whilephlebotomy is unpleasant for most neurotypical children and adults, itis quite traumatic for those with extreme sensory sensitivities who areunable to adequately communicate their fears. Indeed, it is a catch-22for these individuals, in that many of the aberrant behaviors theyexhibit are likely due to an inability to interact, communicate andconnect with others. The potential to provide them with a therapy thatnot only improves upon the core symptoms responsible for thosebehaviors, but to do so without the need for invasive monitoring will goa long way to improve the quality of life for these children.

Results from Study HT 02-121 provide proof of concept that L1-79 appearsto provide benefit in treating the core symptoms of ASD. ASD is definedprimarily as persistent deficits in social communication and socialinteraction as well as restricted and repetitive behavior patterns,interests or activities. Preliminary evidence from multiple independentassessments specifically used to measure both the social and behavioralintensity of these core symptoms demonstrated consistent trends thatwere repeatable across multiple instruments. While the limited size andshort duration of the study precluded any expectation or ability todemonstrate statistically significant improvements in the outcomemeasures used, the agreement between the multiple measures utilized inthis study is very encouraging. In less than one month of treatment, theblinded assessment by the clinicians involved in the study demonstrateda nearly one-point change in the CGI-S compared to baseline. Similarly,patients receiving L1-79 improved by nearly one point from baseline inADOS-2 scores within the same time period.

Previous experience (see Example 5) with open-label administration ofL1-79 in 10 patients with autism clearly demonstrated that L1-79 has thepotential to improve the core symptoms of autism. In Study HT 02-121,multiple independent efficacy measures including the CGI-S, ADOS-2,SRS-2, ABC-C and RBS-R demonstrated consistent improvements in thetarget score social domains affected by ASD despite the short treatmentperiod and small number of patients. Over the 28-day treatment period,L1-79 was safe and well tolerated. At the conclusion of the study, manyparents wanted their children to continue receiving treatment withL1-79. As a result, a roundtable videos were filmed at both clinicalsites with the intent of allowing these parents to speak directly withFDA about the impact of L1-79 on their children with autism.

Example 7 A Randomized, Double-Blind, Placebo-Controlled Adaptive Trialof L1-79 for the Treatment of the Core Deficits in Social-CommunicationFunction and Adolescents and Adults with Autism Spectrum Disorder

The completed Study HT 02-121 (Example 6) was a Phase II safety study ofL1-79 for the treatment of autism. Study HT 02-121 was a randomizeddouble-blind, placebo-controlled two-cohort, 4-week dose-escalationstudy that incorporated 2 open-label treatment groups to assess thesafety and efficacy of L1-79 100 mg and 200 mg TID in male patientsbetween the ages of 13 and 21 years of age with autism. Results fromStudy HT 02-121 provide proof of concept that L1-79 appears to providebenefit in treating the core symptoms of ASD. Preliminary evidence frommultiple independent assessments specifically used to measure both thesocial and behavioral intensity of these core symptoms demonstratedconsistent trends that were repeatable across multiple instruments.While the limited size and short duration of the study precluded anyexpectation or ability to demonstrate statistically significantimprovements in the outcome measures used, the agreement between themultiple measures utilized in this study is very encouraging. Followinga review of the results from Study HT 02-121 the FDA allowed L1-79 tobegin registration trials leading toward a marketing approval andawarded the L1-79 IND 128673 a Fast Track designation.

As a result, the inventors are proposing an adaptive trial approach in aPhase III setting that will allow an expedient yet thorough approach toevaluating L1-79 as a therapy for treating the defining core deficits insocial communication and interaction. Based on the results from Study HT02-121 (Example 6), proposed are two Phase III randomized, double-blind,parallel group, placebo controlled, clinical studies (Study 301 andStudy 302) utilizing adaptive designs in order to evaluate optimalinclusion criteria, sample size and outcome measures during the firstsegment of Study 301 to quantify the safety and efficacy of L1-79administered TID at doses of 200 or 300 mg to subjects with a diagnosisof autism based upon their Autism Diagnostic Observation Schedule-2(ADOS-2) results in a prospectively randomized and double-blind manner.

Most of the clinical experience with L1-79 for the treatment of autismis with doses of 100-200 mg TID. The study of Example 8 (also referredto herein as “Study 301”) seeks to quantify the safety and efficacy ofL1-79 administered TID at doses of 200 or 300 mg to subjects with adiagnosis of autism based upon their Autism Diagnostic ObservationSchedule-2 (ADOS-2) results in a prospectively randomized anddouble-blind manner.

The dosage form is a capsule containing 100 mg ofDL-α-methyl-para-tyrosine capsules (hereafter referred to as “L1-79”).The intended dosing regimen of L1-79 for Phase III clinical studies(Study 301 and Study 302) is 2 or 3 capsules administered orally threetimes daily (TID).

Rationale for Study Design:

ASD is a disorder marked by deficits in social interaction and thepresence of restricted, repetitive patterns of behavior, interests, oractivities during childhood development (Swedo, S. E., Baird, G., Cook,E. H., Happe', F. G., Harris, J. C., Kaufmann, W. E., . . . Wright,Harry, H. (Eds.). (2013). Neurodevelopmental Disorders. In AmericanPsychiatric Association. Diagnostic and statistical manual of mentaldisorders (5th ed.). American Psychiatric Association.). Recentliterature has implicated peripheral and autonomic nervous systeminvolvement in children and adults with ASD (Baker, 2017; Fenning,2017). As a result, electrodermal skin testing may serve as an importantbiomarker for the population of patients more likely to respond totreatments targeting the sympathetic nervous system.

Primary Objective(s):

The primary objectives of the study are to evaluate the efficacy,safety, and tolerability of L1-79 compared to placebo for the treatmentof the core deficits in social communication and interaction inadolescents and adults with autism spectrum disorder (ASD).

Secondary Objective(s):

Secondary objectives of the study include the following:

-   -   1. to evaluate optimal inclusion criteria, sample size, and        outcome measures during the first segment of this adaptive        trial;    -   2. to evaluate the effect of L1-79 compared to placebo on        reducing repetitive and restrictive behaviors in ASD;    -   3. to evaluate the effect of L1-79 compared to placebo on        reducing aberrant behaviors in ASD, including hyperactivity,        agitation, and irritability; and    -   4. to perform additional modeling of pharmacokinetics (PK) and        pharmacodynamics (PD) for L1-79 in a subset of subjects through        timed intermittent sparse sampling.        Study Design:

Eligible participants will be adolescents and adults between the ages of12 and 21 years who meet the Diagnostic and Statistical Manual of MentalDisorders-5th edition (DSM-5) criteria for ASD, based upon clinicianinterview and assessment of ASD symptoms on the ADOS-2, and a ClinicalGlobal Impression of Severity (CGI-S) rating of 4 or greater (moderateor higher).

Subjects will be randomized to placebo, L1-79 200 mg, or L1-79 300 mgthree times daily (TID) groups in a 1:2:2 ratio.

Subjects in the 200 and 300 mg group will be started on 200 mg TID.Subjects in the 300 mg group will titrate up to 300 mg TID after 7 days.

Screening assessments will include the ADOS-2, a review of ASD criteriafrom the DSM-5 and the CGI-S.

An adaptive design will be used with a proposed interim analysis beingused to make decisions on the following criteria; potential endpointsfor the primary outcome, modification of the defined study population,and estimated sample size. The data monitoring committee and independentstatistical group will implement the interim analysis and makerecommendations to the sponsor regarding proposed changes in the studybased on predefined criteria. The sample size recalculation will dependon the primary outcome and potential study populations that will bedetermined at the interim analysis. The sample size recalculation willbe based on blinded data.

A schedule of events is provided in Table 30.

TABLE 30 Schedule of Events Treatment Period Follow-up Screening Visit 1Visit 2 Visit 3 Visit 4 Visit 5 Visit 6 Evaluation Days −30 to −1 Day 0Day 14 = 3 Day 28 = 3 Day 60 = 3 Day 84 = 3 Day 120 = 3 Informed ContextX Inclusion/Exclusion Criteria X Demographics X Medical History¹ X XPhysical Examination X^(2,3) X⁴ X² Vital Signs³ X X X X X X X Urine DrugScreen⁶ X X Pregnancy Test⁷ X X Pharmacokinetics (PK) - 1 hr post dose XPK - Random (known time after dose) X⁸ X⁸ X Laboratory Tests⁹ X X XRandomization X 12-Lead ECG X X X X Drug Administration X X X X DrugAccountability X X X X Adverse Event (AE) Assessment X X X X X XPrior/Concomitant Medication X X X X X X X Assessment ADOS-2¹⁰ X X CGI-SX X X X CGI-C X X X X VABS-3¹¹ X X X X X SRS-2 X X X X X ABC-C X X X X XRBS-R X X X X X PSI X X X SSP X X WASI-2 X DSM-5 criteria for ASD X SkinReactivity Testing (SRT) X X X ABC-C = Aberrant BehaviorChecklist-Community; ADOS-2 = Autism Diagnostic Observation Scale-2; ASD= autism spectrum disorder; CGI-C = Clinical Global Impression ofChange; CGI-S = Clinical Global Impression of Severity; DSM-5 =Diagnostic and Statistical Manual of Mental Disorders-5th edition; PSI =Parenting Stress Index; RBS-R = Repetitive Behavior Scale-Revised; SAS =Spence Anxiety Scale; SSP = Spence Anxiety Scale; SRS-2 = SocialResponsiveness Scale-2; VABS-3 = Vineland Adaptive Behavior Scales - 3rdEdition; WASI-2 - Weschler Abbreviated Scales of Intelligence ¹Includesa review of previous/ongoing medications ²Complete examination,including assessments of the skin, head, eyes, ears, nose, throat, neck,thyroid, lungs, heart, abdomen, lymph nodes, extremities, and bodyweight ³Height will be measured at screening only. ⁴Partial examination,to update findings from the examination performed at screening ⁵Includesrespiratory rate, oral temperature, sitting and standing orthostaticblood pressure and pulse ⁶Includes amphetamines, barbiturates, cocainemetabolites, opiates, benzodiazepines, cannabinoids, and cotinine ⁷Onlyfor females who are post-menarche ⁸Subject will only have random PK donefor one of Visits 2, 3, or 4. ⁹Includes serum chemistry, hematology(including coagulation), and urinalysis ¹⁰ADOS-2 Will be videotaped forpotential use as an additional review for Reciprocal Social Interactionby Blinded reviewer ¹¹Adaptive Behavior Composite domains onlyInclusion Criteria:1. Subjects must be male or female adolescents or adults up to age 212. Subjects must be between the ages of 13 and 21 years of age3. Post menarche females must be on birth control if appropriate.4. Diagnosis of ASD based upon an assessment tool that utilizes theDSM-5 criteria [e.g., Autism Symptom Rating Scale (ASRS), ChildhoodAutism Rating Scale-2 (CARS2), or Autism Diagnostic Interview-Revised(ADI-R)], and confirmed with the ADOS-2, with a CGI-S score of 4 orgreater.5. Subject must be stable on no more than one concomitant medication,and no planned changes in psychosocial interventions during the trial6. Subjects and caregiver must be willing and able to participate in thetesting procedures sufficient to obtain valid scores on the tests usedherein.7. Subjects must have a caregiver who has known the them for over ayear, spends at least 10 hours per week with them, and is willing toaccompany them to each appointment.8. Subjects must, in the opinion of the Investigator, be sufficientlytolerant and capable of complying with the requirements of this trial.For example, patients who will not tolerate blood draws or ECG are notqualified candidates for this study.9. Subjects must be able to swallow capsules10. Subjects and their care givers must be willing to sign informedconsent or to have informed consent provided by their legal guardians orproxies. All subjects <18 years old or those unable to care forthemselves must have the caregiver's consent.

Exclusion Criteria:

1. Sexually active males and females

2. Uncontrolled intercurrent illness including, but not limited to,ongoing or active infection, symptomatic cardio-vascular disease,hepatic disease, renal disease, skeletomuscular disease, HIV, HCVA, HBV,or psychiatric illness/social situations that would limit compliancewith study requirements.3. Any disease that requires treatment with immunosuppressive drugs4. A diagnosis of Fragile-X syndrome, Rett syndrome, or otherneurological disorder that could be the basis for the subjects autisticsymptoms (e.g., congenital or acquired brain injury, brainmalformations, stroke, neurogenetic or metabolic disorder).5. A DSM-5 diagnosis of schizophrenia, schizoaffective disorder, alcoholuse disorder or ADHD, Current or lifetime diagnosis of severepsychiatric disorder (e.g., bipolar disorder, etc.);6. The Presence of any active chronic medical problem including, but notlimited to uncontrolled seizure disorder, heart disease, cancer, asthma,genetic disease, or any disease or syndrome that requires continuousdrug therapy.7. Subjects requiring more than 1 medication for the treatment ofautism, or who have not been weaned to their lowest tolerable dose ofmedication.8. Subjects with any disease that requires treatment withimmunosuppressive drugs.9. The presences of out of range hepatic or renal function tests orother unexplained abnormal laboratory value that is deemed clinicallysignificant by the Investigator.10. Any subject or caregiver who is unwilling or unable to give informedconsent.Study Population:

350 patients are planned.

Test Product, Dose, and Mode of Administration:

D-L alpha-methyl-tyrosine (L1-79) encapsulated as 100 and 200 mg. Twodose arms will be tested as 200 mg TID (one 200 mg dose combined withplacebo) and 300 mg TID (one 200 mg capsule and one 100 mg capsule)

Duration of Treatment:

-   -   12 weeks (84 days)        Efficacy Assessments:

Baseline assessments not included in the measurement of efficacy includethe Weschler Abbreviated Scales of Intelligence (WASI-2) and the SpenceAnxiety Scale (SAS). Electrodermal testing will be performed at baselineto determine if greater autonomic nervous system variability correlateswith response to treatment. (See Study Rationale). The primary outcomemeasure will be the change from baseline at Week 12 on a 5-factorcomposite measure comprising the Socialization (SOC), Communication(COM), and Daily Living Skills (DLS) domains from the Adaptive BehaviorComposite (ABC) of the Vineland Adaptive Behavior Scales—3rd Edition(VABS-3) and the Socialization, Communication and Interaction (SCI) andRestricted Interests and Repetitive Behavior (RRB) subscales of theSocial Responsiveness Scale-2 (SRS-2). The primary secondary outcomemeasure will be based on the change from baseline CGI-S at Week 12.

Additional secondary outcome measures will be assessed in a gatedfashion (to be determined during the interim analysis) for the changefrom baseline scores at Week 12 for:

1. Aberrant Behavior Checklist-Community (ABC-C) SocialWithdrawal/Lethargy and Inappropriate Speech Domains

2. ABC-C, Irritability, Hyperactivity, and Stereotypy Domains

3. Repetitive Behavior Scale-Revised (RBS-R)

4. Parenting Stress Index (PSI) Short Form

5. Sensory Profile-Short Form (SSP)

6. Spence Anxiety Scale (SAS)

7. ADOS-2 Total Score* (ADOS-2 will be videotaped at screening and atend of study for potential use in blinded review of reciprocalinteraction.)

Safety Assessments:

Safety assessments include physical examination, orthostaticmeasurements of blood pressure and pulse, standard hematology andclinical chemistry assessments, concomitant medication use, urinalysis,ECGs, and spontaneously reported adverse events.

PK Assessments:

In order to gain a greater understanding of the PK and PD of L1-79 inpatients with ASD, sparse sampling will be utilized to assist inmodeling PK parameters and comparing the same to data obtained from morethorough studies done in young healthy adults. Sparse sampling willlimit the number of phlebotomy procedures, an important consideration inchildren with ASD who are typically much more traumatized by blooddrawing procedures than peers without ASD.

Statistical Methods:

The adaptive design will assess three components of the study; theoutcome variable, a potential modification of the inclusion criteria,and a sample size recalculation based on the decisions made at the timeof the interim analysis. The modification of the inclusion criteria willbe based on an evaluation of potential endophenotypes for ASD ofbaseline severity of the following characteristics known to directlyinfluence treatment and intervention strategies for people with ASD: 1)Severity of anxiety based on assessment by the Spence Anxiety Scale, IQas assessed by the Weschler Abbreviated Scale of Intelligence anddisruptive behavior symptom severity, based on the ABC-C irritabilityand hyperactivity subscales. In addition, skin reactivity testing todetermine autonomic sensitivity will be used as an exploratory biomarkerfor responder phenotype (See Study Rationale for justification for thesesubgroups). The choice of primary outcome(s) will be based on aconditional power analysis of the 5-factor composite measure comprisingthe Socialization (SOC), Communication (COM), and Daily Living Skills(DLS) domains from the Adaptive Behavior Composite (ABC) of the VinelandAdaptive Behavior Scales—3rd Edition (VABS-3) and the Socialization,Communication and Interaction (SCI) and Restricted Interests andRepetitive Behavior (RRB) subscales of the Social Responsiveness Scale-2(SRS-2). After selection of the outcomes and inclusion criteria, thesample size will be recalculated based on blinded data.

It will be appreciated by those skilled in the art that changes could bemade to the embodiments described above without departing from the broadinventive concept thereof. It is understood, therefore, that thisinvention is not limited to the particular embodiments disclosed, but itis intended to cover modifications that are within the spirit and scopeof the invention, as defined by the appended claims.

What is claimed is:
 1. A method for treating an autism in a subject inneed thereof, the method comprising administering to said subject acomposition comprising a therapeutically effective amount ofα-methyl-DL-tyrosine, and a pharmaceutically acceptable carrier.
 2. Themethod of claim 1, wherein the composition comprisesα-methyl-DL-tyrosine in an amount ranging from about 50 mg (w/w) toabout 1000 mg (w/w).
 3. The method of claim 1, wherein the compositioncomprises α-methyl-DL-tyrosine in an amount of about 100 mg (w/w/). 4.The method of claim 1, wherein the composition comprisesα-methyl-DL-tyrosine in an amount of about 200 mg (w/w/).
 5. The methodof claim 1, wherein the composition comprises α-methyl-DL-tyrosine in anamount of about 250 mg (w/w/).
 6. The method of claim 1, wherein thecomposition comprises α-methyl-DL-tyrosine in an amount of about 300 mg(w/w/).
 7. The method of claim 1, wherein the composition isadministered in a plurality of divided doses.
 8. The method of claim 1,wherein the composition is administered in a plurality of divided doses.9. The method of claim 1, wherein the composition is administered as asingle sustained release daily dose.
 10. The method of claim 1, whereinthe composition is administered as a chewable dosage form for pediatricpatients.
 11. The method of claim 1, wherein the composition isadministered two times per day.
 12. The method of claim 1, wherein thecomposition is administered three times per day.
 13. The method of claim1, further comprising administering to said subject a therapeuticallyeffective amount of a central nervous system (CNS) agent.
 14. The methodof claim 13, wherein said agent is Y-aminobutyric acid (GABA).
 15. Themethod of claim 13, wherein said agent is beta-adrenergic agonist. 16.The method of claim 1, wherein the composition is administered orally.17. The method of claim 16, wherein the composition is in a form of acapsule or a tablet.
 18. The method of claim 1, wherein said subject isa human patient between 3 years of age and 21 years of age.
 19. Themethod of claim 1, wherein said subject is an adult human patient.